掲載種別：研究論文（学術雑誌）  

DOI： 10.52794/hujpharm.1354419

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

A Cucurbita phloem exudate lectin (CPL) from summer squash (Cucurbita pepo) fruits was isolated and its sugar-binding properties and biological activities were studied. The lectin was purified by affinity chromatography and the hemagglutination assay method was used to determine its pH, heat stability, metal-dependency and sugar specificity. Antimicrobial and anticancer activities were also studied by disc diffusion assays and in vivo and in vitro methods. The molecular weight of CPL was 30 ± 1 KDa and it was stable at different pH (5.0 to 9.0) and temperatures (30 to 60 °C). CPL recovered its hemagglutination activity in the presence of Ca2+. 4-nitrophenyl-α-D-glucopyranoside, lactose, rhamnose and N-acetyl-D-glucosamine strongly inhibited the activity. With an LC50 value of 265 µg/mL, CPL was moderately toxic and exhibited bacteriostatic, bactericidal and antibiofilm activities against different pathogenic bacteria. It also exhibited marked antifungal activity against Aspergillus niger and agglutinated A. flavus spores. In vivo antiproliferative activity against Ehrlich ascites carcinoma (EAC) cells in Swiss albino mice was observed when CPL exerted 36.44% and 66.66% growth inhibition at doses of 3.0 mg/kg/day and 6.0 mg/kg/day, respectively. A 12-day treatment by CPL could reverse their RBC and WBC counts as well as restore the hemoglobin percentage to normal levels. The MTT assay of CPL performed against human breast (MCF-7) and lung (A-549) cancer cell lines showed 29.53% and 18.30% of inhibitory activity at concentrations of 128 and 256 µg/mL, respectively.

DOI： 10.3390/molecules29112531

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

This research describes the synthesis by an environmentally-friendly method, microwave irradiation, development and analysis of three novel and one previously identified Schiff base derivative as a potential inhibitor of bovine xanthine oxidase (BXO), a key enzyme implicated in the progression of gout. Meticulous experimentation revealed that these compounds (10, 9, 4, and 7) have noteworthy inhibitory effects on BXO, with IC50 values ranging from 149.56 µM to 263.60 µM, indicating their good efficacy compared to that of the standard control. The validation of these results was further enhanced through comprehensive in silico studies, which revealed the pivotal interactions between the inhibitors and the catalytic sites of BXO, with a particular emphasis on the imine group (-C = N-) functionalities. Intriguingly, the compounds exhibiting the highest inhibition rates also showcase advantageous ADMET profiles, alongside encouraging initial assessments via PASS, hinting at their broad-spectrum potential. The implications of these findings are profound, suggesting that these Schiff base derivatives not only offer a new vantage point for the inhibition of BXO but also hold considerable promise as innovative therapeutic agents in the management and treatment of gout, marking a significant leap forward in the quest for more effective gout interventions.

DOI： 10.1016/j.jsps.2024.102062

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Nucleoside analogs have been widely used as antiviral, antitumor, and antiparasitic agents due to their ability to inhibit nucleic acid synthesis. Adenosine, cytidine, guanosine, thymidine and uridine analogs such as didanosine, vidarabine, remdesivir, gemcitabine, lamivudine, acyclovir, abacavir, zidovusine, stavudine, and idoxuridine showed remarkable anticancer and antiviral activities. In our previously published articles, our main intention was to develop newer generation nucleoside analogs with acylation-induced modification of the hydroxyl group and showcase their biological potencies. In the process of developing nucleoside analogs, in silico studies play an important role and provide a scientific background for biological data. Molecular interactions between drugs and receptors followed by assessment of their stability in physiological environments, help to optimize the drug development process and minimize the burden of unwanted synthesis. Computational approaches, such as DFT, FMO, MEP, ADMET prediction, PASS prediction, POM analysis, molecular docking, and molecular dynamics simulation, are the most popular tools to culminate all preclinical study data and deliver a molecule with maximum bioactivity and minimum toxicity. Although clinical drug trials are crucial for providing dosage recommendations, they can only indirectly provide mechanistic information through researchers for pathological, physiological, and pharmacological determinants. As a result, in silico approaches are increasingly used in drug discovery and development to provide mechanistic information of clinical value. This article portrays the current status of these methods and highlights some remarkable contributions to the development of nucleoside analogs with optimized bioactivity.

DOI： 10.2174/0113895575258033231024073521

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Elsevier BV  

DOI： 10.1016/j.ijbiomac.2023.127628

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Elsevier BV  

DOI： 10.1016/j.jsps.2023.101804

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担当区分：最終著者   掲載種別：研究論文（学術雑誌）  

DOI： 10.3390/md21120614

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担当区分：最終著者   記述言語：英語  

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担当区分：最終著者,　責任著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Association of Carbohydrate Chemists and Technologists, India (ACCTI www.accti.in)  

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その他リンク： https://www.trendscarbo.com/aboutjournal.php

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.3390/molecules28010219

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担当区分：責任著者   記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.3390/md20100613

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

PubMed

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担当区分：最終著者   記述言語：英語   出版者・発行元：Cold Spring Harbor Laboratory Press  

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その他リンク： https://www.ncbi.nlm.nih.gov/books/NBK579918/toc/?report=reader

記述言語：英語   掲載種別：研究論文（学術雑誌）  

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掲載種別：研究論文（学術雑誌）  

Carbohydrate esters are significant in medicinal chemistry because of their efficacy for the synthesis of biologically active drugs. In the present study, methyl β-D-galactopyranoside (MGP) was treated with various acyl halides to produce 6-O-acyl MGP esters by direct acylation method with an excellent yield. To obtain newer products for antimicrobial assessment studies, the 6-O-MGP esters were further modified into 2,3,4-tri-O-acyl MGP esters containing a wide variety of functionalities in a single molecular framework. The chemical structures of the newly synthesized compounds were elucidated by analyzing their physicochemical, elemental, and spectroscopic data. In vitro antimicrobial testing against five bacteria and two fungi and the prediction of activity spectra for substances (PASS) revealed that these MGP estes have promising antifungal functionality compared to their antibacterial activities. The antimicrobial tests demonstrated that the compounds 3 and 10 were the most potent against Bacillus subtilis and Escherichia coli strains, with the minimum inhibitory concentration (MIC) values ranging from 0.352 ± 0.02 to 0.703 ± 0.01 mg/ml and minimum bactericidal concentration (MBC) values ranging from 0.704 ± 0.02 to 1.408 ± 0.04 mg/ml. Density functional theory (DFT) at the B3LYP/3-21G level of theory was employed to enumerate, frontier orbital energy, enthalpy, free energy, electronic energy, MEP, dipole moment which evaluated the effect of certain groups (aliphatic and aromatic) on drug properties. They discovered that all esters were more thermodynamically stable than the parent molecule. Molecular docking is performed using AutoDock Vina to determine the binding affinities and interactions between the MGP esters and the SARS-CoV-2 main protease. The modified esters strongly interact with the prime Cys145, His41, MET165, GLY143, THR26, and ASN142 residues. The MGP esters' shape and ability to form multiple electrostatic and hydrogen bonds with the active site match other minor-groove binders' binding modes. The molecular dynamics simulation validates the molecular docking results. The pharmacokinetic characterization of the optimized inhibitor demonstrates that these MGP esters appear to be safer inhibitors and a combination of in silico ADMET (absorption, distribution, metabolism, excretion, and toxicity) prediction and drug-likeness had promising results due to their improved kinetic properties. Structure activity relationships (SAR) study including in vitro and silico results revealed that the acyl chain, palmitoyl (C16) and 4-chlorobenzoyl (4.ClC₆H₄CO-) in combination with sugar were found the most potential activates against human and fungal pathogens. After all, our comprehensive computational and statistical analysis shows that these selected MGP esters can be used as potential inhibitors against the SARS-CoV-2.

DOI： 10.1007/s10719-021-10039-3

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担当区分：責任著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：MDPI AG  

DOI： 10.3390/molecules26227016

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担当区分：責任著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：MDPI  

DOI： 10.3390/molecules26164799

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.3390/md19070394

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.29356/jmcs.v65i2.1464

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掲載種別：研究論文（学術雑誌）   出版者・発行元：Pensoft Publishers  

Nucleoside derivatives are important therapeutic drugs and are the focal point in the ongoing search for novel, more potent drug targets. In this study, a new series of pyrimidine nucleoside i.e., uridine (<bold>1</bold>) derivatives were synthesized via direct method and evaluated for their antimicrobial potential activity. The title compound uridine (<bold>1</bold>) was treated with triphenylmethyl chloride in pyridine to give the 5´-<italic>O</italic>-(triphenylmethyl)uridine derivative (<bold>2</bold>), which was subsequently derivatized to create a series of 2´,3´-di-<italic>O</italic>-acyl analogs containing a wide variety of functionalities in a single molecular framework. <italic>In vitro</italic> antimicrobial functionality tests were determined against both human and plant pathogens by disc diffusion and food poisoned techniques. The chemical structures of the synthesized compounds were confirmed on the basis of their spectral, analytical, physicochemical data. The antimicrobial results indicated that the synthesized derivatives exhibited moderate to good antibacterial and antifungal activity; in particular, they were found to be more effective against fungal phytopathogens than against human bacterial strains. Compounds <bold>7</bold>, <bold>9</bold>, and <bold>14</bold> were of particular interest as they exhibited noteworthy antifungal and antibacterial properties. <italic>In vitro</italic> MTT assays revealed that compound <bold>9</bold> was effective against Ehrlich’s ascites carcinoma (EAC) cells, resulting in 7.12% and 1.34% cell growth inhibition at concentrations of 200 and 6.25 µg/ml, respectively. The IC₅₀ value for compound <bold>9</bold> was rather high and found to be 1956.25 µg/ml. Structure-activity relationship (SAR) studies were also conducted to predict structural and pharmacokinetic properties. The findings of this study indicate that the different uridine derivatives are potentially useful antimicrobial agents for the advancement of future pharmaceutical research.

DOI： 10.3897/pharmacia.68.e56543

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その他リンク： https://pharmacia.pensoft.net/article/56543/download/xml/

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.2478/auoc-2021-0002

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Nucleoside analogs contribute in pharmaceutical and clinical fields as medicinal agents and approved drugs. This work focused to investigate the antimicrobial, anticancer activities, and structure-activity relationship (SAR) of cytidine and its analogs with computational studies. Microdilution was used to determine the antimicrobial activity, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) of the modified analogs against human and phytopathogenic strains. Compounds (7), (10), and (14) were the most potent against Escherichia coli and Salmonella abony strains with MIC and MBC values from 0.316 ± 0.02 to 2.50 ± 0.03 and 0.625 ± 0.04 to 5.01 ± 0.06 mg/ml, respectively. The highest inhibitory activity was observed against gram-positive bacteria. Numerous analogs (10), (13), (14), and (15) exhibited good activity against the tested fungi Aspergillus niger and Aspergillus flavus. Anticancer activity of the cytidine analogs was examined through MTT colorimetric assay against Ehrlich's ascites carcinoma (EAC) tumor cells whereas compound 6 showed the maximum antiproliferative activity with an IC50 value of 1168.97 µg/ml. To rationalize this observation, their quantum mechanical and molecular docking studies have been performed against urate oxidase of A. flavus 1R51 to investigate the binding mode, binding affinity, and non-bonding interactions. It was observed that most of the analogs exhibited better binding properties than the parent drug. In silico ADMET prediction was attained to evaluate the drug-likeness properties that revealed the improved pharmacokinetic profile with lower acute oral toxicity of cytidine analogs. Based on the in vitro and in silico analysis, this exploration can be useful to develop promising cytidine-based antimicrobial drug(s). Supplementary Information: The online version contains supplementary material available at 10.1007/s40203-021-00102-0.

DOI： 10.1007/s40203-021-00102-0

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Springer Nature Ltd  

添付ファイル： Kamata et al (2020) Sci Rep 10_22102.pdf

DOI： 10.1038/s41598-020-78926-7

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出版者・発行元：Book Publisher International (a part of {SCIENCEDOMAIN} International)  

DOI： 10.9734/bpi/cpcs/v3

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

添付ファイル： Sugawara et al (2020) Glycobiology.pdf

DOI： 10.1093/glycob/cwaa029

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その他リンク： http://academic.oup.com/glycob/article-pdf/30/10/802/33807038/cwaa029.pdf

担当区分：責任著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Wiley  

A 15-kDa lectin, termed SeviL, was isolated from Mytilisepta virgata (purplish bifurcate mussel). SeviL forms a noncovalent dimer that binds strongly to ganglio-series GM1b oligosaccharide (Neu5Acɑ2-3Galβ1-3GalNAcβ1-4Galβ1-4Glc) and its precursor, asialo-GM1 (Galβ1-3GalNAcβ1-4Galβ1-4Glc). SeviL also interacts weakly with the glycan moiety of SSEA-4 hexaose (Neu5Acα2-3Galβ1-3GalNAcβ1-3Galα1-4Galβ1-4Glc). A partial protein sequence of the lectin was determined by mass spectrometry, and the complete sequence was identified from transcriptomic analysis. SeviL, consisting of 129 amino acids, was classified as an R(icin B)-type lectin, based on the presence of the QxW motif characteristic of this fold. SeviL mRNA is highly expressed in gills and, in particular, mantle rim tissues. Orthologue sequences were identified in other species of the family Mytilidae, including Mytilus galloprovincialis, from which lectin MytiLec-1 was isolated and characterized in our previous studies. Thus, mytilid species contain lectins belonging to at least two distinct families (R-type lectins and mytilectins) that have a common β-trefoil fold structure but differing glycan-binding specificities. SeviL displayed notable cytotoxic (apoptotic) effects against various cultured cell lines (human breast, ovarian, and colonic cancer; dog kidney) that possess asialo-GM1 oligosaccharide at the cell surface. This cytotoxic effect was inhibited by the presence of anti-asialo-GM1 oligosaccharide antibodies. With HeLa ovarian cancer cells, SeviL showed dose- and time-dependent activation of kinase MKK3/6, p38 MAPK, and caspase-3/9. The transduction pathways activated by SeviL via the glycosphingolipid oligosaccharide were triggered apoptosis. DATABASE: Nucleotide sequence data have been deposited in the GenBank database under accession numbers MK434191, MK434192, MK434193, MK434194, MK434195, MK434196, MK434197, MK434198, MK434199, MK434200, and MK434201.

添付ファイル： Fuii et al (2020) FEBS J 287 2612-2630.pdf

DOI： 10.1111/febs.15154

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その他リンク： https://onlinelibrary.wiley.com/doi/full-xml/10.1111/febs.15154

担当区分：最終著者,　責任著者   記述言語：英語   掲載種別：論文集(書籍)内論文   出版者・発行元：Springer US  

DOI： 10.1007/978-1-0716-0430-4_21

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.25177/jccmm.4.4.ra.10663

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出版者・発行元：MDPI  

DOI： 10.3390/books978-3-03921-821-9

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：MDPI AG  

MytiLec-1, a 17 kDa lectin with β-trefoil folding that was isolated from the Mediterranean mussel (Mytilus galloprovincialis) bound to the disaccharide melibiose, Galα(1,6) Glc, and the trisaccharide globotriose, Galα(1,4) Galβ(1,4) Glc. Toxicity of the lectin was found to be low with an LC50 value of 384.53 μg/mL, determined using the Artemia nauplii lethality assay. A fluorescence assay was carried out to evaluate the glycan-dependent binding of MytiLec-1 to Artemia nauplii. The lectin strongly agglutinated Ehrlich ascites carcinoma (EAC) cells cultured in vivo in Swiss albino mice. When injected intraperitoneally to the mice at doses of 1.0 mg/kg/day and 2.0 mg/kg/day for five consecutive days, MytiLec-1 inhibited 27.62% and 48.57% of cancer cell growth, respectively. Antiproliferative activity of the lectin against U937 and HeLa cells was studied by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in vitro in RPMI-1640 medium. MytiLec-1 internalized into U937 cells and 50 μg/mL of the lectin inhibited their growth of to 62.70% whereas 53.59% cell growth inhibition was observed against EAC cells when incubated for 24 h. Cell morphological study and expression of apoptosis-related genes (p53, Bax, Bcl-X, and NF-κB) showed that the lectin possibly triggered apoptosis in these cells.

DOI： 10.3390/md17090502

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担当区分：最終著者   記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.marpolbul.2019.02.061

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担当区分：責任著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Elsevier {BV}  

DOI： 10.1016/j.ijbiomac.2018.11.222

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担当区分：責任著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：MDPI AG  

We identified a lectin (carbohydrate-binding protein) belonging to the complement 1q(C1q) family in the feather star Anneissia japonica (a crinoid pertaining to the phylum Echinodermata). The combination of Edman degradation and bioinformatics sequence analysis characterized the primary structure of this novel lectin, named OXYL, as a secreted 158 amino acid-long globular head (sgh)C1q domain containing (C1qDC) protein. Comparative genomics analyses revealed that OXYL pertains to a family of intronless genes found with several paralogous copies in different crinoid species. Immunohistochemistry assays identified the tissues surrounding coelomic cavities and the arms as the main sites of production of OXYL. Glycan array confirmed that this lectin could quantitatively bind to type-2 N-acetyllactosamine (LacNAc: Galβ1-4GlcNAc), but not to type-1 LacNAc (Galβ1-3GlcNAc). Although OXYL displayed agglutinating activity towards Pseudomonas aeruginosa, it had no effect on bacterial growth. On the other hand, it showed a significant anti-biofilm activity. We provide evidence that OXYL can adhere to the surface of human cancer cell lines BT-474, MCF-7, and T47D, with no cytotoxic effect. In BT-474 cells, OXYL led to a moderate activation of the p38 kinase in the MAPK signaling pathway, without affecting the activity of caspase-3. Bacterial agglutination, anti-biofilm activity, cell adhesion, and p38 activation were all suppressed by co-presence of LacNAc. This is the first report on a type-2 LacNAc-specific lectin characterized by a C1q structural fold.

DOI： 10.3390/md17020136

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Acta Pharmaceutica Sciencia  

DOI： 10.23893/1307-2080.aps.05704

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担当区分：責任著者   記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.4052/tigg.1717.1j

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担当区分：責任著者   記述言語：日本語   掲載種別：研究論文（学術雑誌）  

添付ファイル： Fujii et al TIGG 30 177J155-188_1717.1J.pdf

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：BIOMED CENTRAL LTD  

Background: Mytilisepta virgata is a marine mussel commonly found along the coasts of Japan. Although this species has been the subject of occasional studies concerning its ecological role, growth and reproduction, it has been so far almost completely neglected from a genetic and molecular point of view. In the present study we present a high quality de novo assembled transcriptome of the Japanese purplish mussel, which represents the first publicly available collection of expressed sequences for this species.
Results: The assembled transcriptome comprises almost 50,000 contigs, with a N50 statistics of similar to 1 kilobase and a high estimated completeness based on the rate of BUSCOs identified, standing as one of the most exhaustive sequence resources available for mytiloid bivalves to date. Overall this data, accompanied by gene expression profiles from gills, digestive gland, mantle rim, foot and posterior adductor muscle, presents an accurate snapshot of the great functional specialization of these five tissues in adult mussels.
Conclusions: We highlight that one of the most striking features of the M. virgata transcriptome is the high abundance and diversification of lectin-like transcripts, which pertain to different gene families and appear to be expressed in particular in the digestive gland and in the gills. Therefore, these two tissues might be selected as preferential targets for the isolation of molecules with interesting carbohydrate-binding properties. In addition, by molecular phylogenomics, we provide solid evidence in support of the classification of M. virgata within the Brachidontinae subfamily. This result is in agreement with the previously proposed hypothesis that the morphological features traditionally used to group Mytilisepta spp. and Septifer spp. within the same clade are inappropriate due to homoplasy.

DOI： 10.1186/s12864-017-4012-z

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1038/s41598-017-06332-7

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：WILEY  

Background Botrocetin-2 (Bot2) is a botrocetin-like protein composed of and subunits that have been cloned from the snake Bothrops jararaca. Bot2 binds specifically to von Willebrand factor (VWF), and the complex induces glycoprotein (GP) Ib-dependent platelet agglutination.
Objectives To exploit Bot2's VWF-binding capacity in order to attempt to create a mutant Bot2 that binds to VWF but inhibits platelet agglutination.
Methods and Results Several point mutations were introduced into Bot2 cDNA, and the recombinant protein (recombinant Bot2 [rBot2]) was purified on an anti-botrocetin column. The mutant rBot2 with either Ala at Asp70 in the subunit (Asp70Ala), or Arg115Ala and Lys117Ala, showed reduced platelet agglutination-inducing activity. rBot2 with Asp70Ala showed little binding activity towards immobilized VWF on an ELISA plate, whereas rBot2 with Arg115Ala/Lys117Ala showed reduced binding activity towards GPIb (glycocalicin) after forming a complex with VWF. rBot2 point-mutated to oppositely charged Glu at both Arg115 and Lys117 showed normal binding activity towards VWF but no platelet-agglutinating activity. Furthermore, this doubly mutated protein inhibited ristocetin-induced or high shear stress-induced platelet aggregation, and restrained thrombus formation under flow conditions.
Conclusions Asp70 in the subunit of botrocetin is important for VWF binding, and Arg115 and Lys117 in the subunit are essential for interaction with GPIb. Doubly mutated rBot2, with Arg115Glu and Lys117Glu, repels GPIb and might have potential as an antithrombotic reagent that specifically blocks VWF function. This is the first report on an artificial botrocetin that can inhibit the VWF-GPIb interaction.

DOI： 10.1111/jth.13617

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SPRINGER  

An N-acetyl sugar-binding lectin (termed iNoL) displaying cytotoxic activity against human cancer cells was isolated from the slipper lobster Ibacus novemdentatus (family Scyllaridae). iNoL recognized monosaccharides containing N-acetyl group, and glycoproteins (e.g., BSM) containing oligosaccharides with N-acetyl sugar. iNoL was composed of five subunits (330, 260, 200, 140, and 30 kDa), which in turn consisted of 70-, 40-, and 30-kDa polypeptides held together by disulfide bonds. Electron microscopic observations and gel permeation chromatography indicated that iNoL was a huge (500-kDa) molecule and had a polygonal structure under physiological conditions. iNoL displayed cytotoxic (apoptotic) effects against human cancer cell lines MCF7 and T47D (breast), HeLa (ovarian), and Caco2 (colonic), through incorporation (internalization) into cells. The lectin was transported into lysosomes via endosomes. Its cytotoxic effect and incorporation into cells were inhibited by the co-presence of N-acetyl-D-mannosamine (ManNAc). Treatment of HeLa cells with iNoL resulted in DNA fragmentation and chromatin condensation, through activation of caspase-9 and -3. In summary, the novel crustacean lectin iNoL is incorporated into mammalian cancer cells through glycoconjugate interaction, and has cytotoxic (apoptotic) effects.

添付ファイル： Fujii et al (2017) Glycoconj J 34 85-94.pdf

DOI： 10.1007/s10719-016-9731-x

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SPRINGER  

Silurus asotus egg lectin (SAL), an alpha-galactoside-binding protein isolated from the eggs of catfish, is a member of the rhamnose-binding lectin family that binds to Gb3 glycan (Gal alpha 1-4Gal beta 1-4Glc). We have previously demonstrated that SAL reduces the proliferation of Gb3-expressing Burkitt's lymphoma Raji cells and confirm here that it does not reduce their viability, indicating that unlike other lectins, it is not cytotoxic. The aim of this study was to determine the signal transduction mechanism(s) underlying this novel SAL/Gb3 binding-mediated effect profile. SAL/Gb3 interaction arrested the cell cycle through increasing the G(0/1) phase population of Raji cells. SAL suppressed the transcription of cell cycle-related factors such as c-MYC, cyclin D3, and cyclin-dependent protein kinase (CDK)-4. Conversely, the CDK inhibitors p21 and p27 were elevated by treatment with SAL. In particular, the production of p27 in response to SAL treatment increased steadily, whereas p21 production was maximal at 12 h and lower at 24 h. Activation of Ras-MEK-ERK pathway led to an increase in expression of p21. Notably, treatment of Raji cells with anti-Gb3 mAb alone did not produce the above effects. Taken together, our findings suggest that Gb3 on the Raji cell surface interacts with SAL to trigger sequential GDP-Ras phosphorylation, Ras-MEK-ERK pathway activation, p21 production, and cell cycle arrest at the G(0/1) phase.

添付ファイル： Sugawara et al (2017) Glycoconj J 34 127-138.pdf

DOI： 10.1007/s10719-016-9739-2

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：WILEY-BLACKWELL  

To detect metalloproteinase-7 (MMP7), zymography is conducted using a casein substrate and conventional CBB stain. It has disadvantages because it is time consuming and has low sensitivity. Previously, a sensitive method to detect MMP7 up to 30 pg was reported, however it required special substrates and complicated handlings. RAMA casein zymography described herein is rapid, sensitive, and reproducible. By applying high-sensitivity staining with low substrate conditions, the staining process is completed within 1 h and sensitivity was increased 100-fold. The method can detect 10 pg MMP7 by using commercially available casein without complicated handlings. Moreover, it increases detection sensitivity for trypsin.

DOI： 10.1002/elps.201600346

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記述言語：英語  

添付ファイル： Ch33 Hasan et al Marine Glycobiology Principles and Applications KIM 2016 CRC.pdf

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記述言語：英語  

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：NATURE PUBLISHING GROUP  

MytiLec is a lectin, isolated from bivalves, with cytotoxic activity against cancer cell lines that express globotriaosyl ceramide, Gal alpha(1,4)Gal beta(1,4) Glc alpha 1-Cer, on the cell surface. Functional analysis shows that the protein binds to the disaccharide melibiose, Gal alpha(1,6)Glc, and the trisaccharide globotriose, Gal alpha(1,4)Gal beta(1,4)Glc. Recombinant MytiLec expressed in bacteria showed the same haemagglutinating and cytotoxic activity against Burkitt's lymphoma (Raji) cells as the native form. The crystal structure has been determined to atomic resolution, in the presence and absence of ligands, showing the protein to be a member of the beta-trefoil family, but with a mode of ligand binding unique to a small group of related trefoil lectins. Each of the three pseudo-equivalent binding sites within the monomer shows ligand binding, and the protein forms a tight dimer in solution. An engineered monomer mutant lost all cytotoxic activity against Raji cells, but retained some haemagglutination activity, showing that the quaternary structure of the protein is important for its cellular effects.

DOI： 10.1038/srep28344

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：MDPI AG  

MytiLec is an -d-galactose-binding lectin with a unique primary structure isolated from the Mediterranean mussel (Mytilus galloprovincialis). The lectin adopts a -trefoil fold that is also found in the B-sub-unit of ricin and other ricin-type (R-type) lectins. We are introducing MytiLec(-1) and its two variants (MytiLec-2 and -3), which both possess an additional pore-forming aerolysin-like domain, as members of a novel multi-genic mytilectin family in bivalve mollusks. Based on the full length mRNA sequence (911 bps), it was possible to elucidate the coding sequence of MytiLec-1, which displays an extended open reading frame (ORF) at the 5 end of the sequence, confirmed both at the mRNA and at the genomic DNA sequence level. While this extension could potentially produce a polypeptide significantly longer than previously reported, this has not been confirmed yet at the protein level. MytiLec-1 was revealed to be encoded by a gene consisting of two exons and a single intron. The first exon comprised the 5UTR and the initial ATG codon and it was possible to detect a putative promoter region immediately ahead of the transcription start site in the MytiLec-1 genomic locus. The remaining part of the MytiLec-1 coding sequence (including the three sub-domains, the 3UTR and the poly-A signal) was included in the second exon. The bacteriostatic activity of MytiLec-1 was determined by the agglutination of both Gram-positive and Gram-negative bacteria, which was reversed by the co-presence of -galactoside. Altogether, these data support the classification of MytiLec-1 as a member of the novel mytilectin family and suggest that this lectin may play an important role as a pattern recognition receptor in the innate immunity of mussels.

DOI： 10.3390/md14050092

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE BV  

Chitinases are a group of enzymes that show differences in their molecular structure, substrate specificity, and catalytic mechanism and widely found in organisms like bacteria, yeasts, fungi, arthropods actinomycetes, plants and humans. A novel chitinase enzyme (designated as TDSC) was purified from Trichosanthes dioica seed with a molecular mass of 39 +/- 1 kDa in the presence and absence of beta-mercaptoethanol. The enzyme was a glycoprotein in nature containing 8% neutral sugar. The N-terminal sequence was determined to be EINGGGA which did not match with other proteins. Amino acid analysis performed by LC-MS revealed that the protein was rich in leucine. The enzyme was stable at a wide range of pH (5.0-11.0) and temperature (30-90 degrees C). Chitinase activity was little bit inhibited in the presence of chelating agent EDTA (ethylenediaminetetraaceticacid), urea and Ca2+. A strong fluorescence quenching effect was found when dithiothreitol and sodium dodecyl sulfate were added to the enzyme. TDSC showed antifungal activity against Aspergillus niger and Trichoderma sp. as tested by MU assay and disc diffusion method. (C) 2015 Elsevier B.V. All rights reserved.

添付ファイル： Kabir et al (2016) Int J Biol Macromol 84 62-68.pdf

DOI： 10.1016/j.ijbiomac.2015.12.006

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

添付ファイル： Kamitori et al (2016) Regenerative Therapy 3 11-14.pdf

DOI： 10.1016/j.reth.2016.01.006

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

添付ファイル： Kawsar et al (2016) Int Lett Chem Phys Astro 64 95-105.pdf

DOI： 10.18052/www.scipress.com/ilcpa.64.95

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Growing Science  

添付ファイル： Chowdhury et al (2016) Curr Chem Lett 5 83-92.pdf

DOI： 10.5267/j.ccl.2015.12.001

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：{MDPI} {AG}  

DOI： 10.3390/molecules21010129

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Scientific Research Publishing, Inc.  

添付ファイル： Kawsar et al (2015) Int J Org Chem USA 5(4) 232-245.pdf

DOI： 10.4236/ijoc.2015.54023

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

添付ファイル： Hasan et al (2015) Mar Drugs 13(12)7377-7389.pdf

DOI： 10.3390/md13127071

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：WILEY-BLACKWELL  

Toward the development of ecotoxicology methods to investigate microbial markers of impacts of hydrocarbon processing activities, DNA adductomic analyses were conducted on a sphingomonad soil bacterium. From growing cells that were exposed or unexposed to acrolein, a commonly used biocide in hydraulic fracturing processes, DNA was extracted, digested to 2'-deoxynucleosides and analyzed by liquid chromatography-positive ionization electrospray-tandem mass spectrometry in selected reaction monitoring mode transmitting the [M + H](+) &gt; [M + H - 116](+) transition over 100 transitions. Overall data shown as DNA adductome maps revealed numerous putative DNA adducts under both conditions with some occurring specifically for each condition. Adductomic analyses of triplicate samples indicated that elevated levels of some targeted putative adducts occurred in exposed cells. Two exposure-specific adducts were identified in exposed cells as 3-(2'-deoxyribosyl)-5,6,7,8-tetrahydro-6-hydroxy- (and 8-hydroxy-)pyrimido[1,2-a]-purine-(3H)-one (6- and 8-hydroxy-PdG) following synthesis of authentic standards of these compounds and subsequent analyses. A time course experiment showed that 6- and 8-hydroxy-PdG were detected in bacterial DNA within 30 min of acrolein exposure but were not detected in unexposed cells. This work demonstrated the first application of DNA adductomics to examine DNA damage in a bacterium and sets a foundation for future work.

DOI： 10.1002/mbo3.283

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

添付ファイル： Kawsar et al (2015) Int Lett Chem Phys Astro 58 122-136.pdf

DOI： 10.18052/www.scipress.com/ilcpa.58.122

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SOC GENERAL MICROBIOLOGY  

A Gram-stain-negative, yellow, rod-shaped bacterium, designated strain KK22(T), was isolated from a microbial consortium that grew on diesel fuel originally recovered from cattle pasture soil. Strain KK22(T) has been studied for its ability to biotransform high molecular weight polycyclic aromatic hydrocarbons. On the basis of 16S rRNA gene sequence phylogeny, strain KK22(T) was affiliated with the genus Sphingobium in the phylum Proteobacteria and was most closely related to Sphingobium fuliginis TKPT (99.8 %) and less closely related to Sphingobium quisquiliarum P25(T) (97.5 %). Results of DNA DNA hybridization (DDH) revealed relatedness values between strain KK22(T) and strain TKPT and between strain KK22(T) and strain P25(T) of 21 +/- 4 % (reciprocal hybridization, 27 +/- 2 %) and 15 +/- 2 % (reciprocal hybridization, 17 +/- 1 %), respectively. Chemotaxonomic analyses of strain KK22(T) showed that the major respiratory quinone was ubiquinone Q-10, that the polar lipid profile consisted of phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, phosphatidyl-N-methylethylethanolamine and sphingoglycolipid, and that C-18 : 1 omega 7c and C-14 :0 2-OH were the main fatty acid and hydroxylated fatty acids, respectively. This strain was unable to reduce nitrate and the genomic DNA G+C content was 64.7 mol%. Based upon the results of the DDH analyses, the fact that strain KK22(T) was motile, and its biochemical and physiological characteristics, strain KK22(T) could be separated from recognized species of the genus Sphingobium. We conclude that strain KK22(T) represents a novel species of this genus for which the name Sphingobium barthaii sp. nov. is proposed; the type strain is KK22(T) (=DSM 29313(T)=JCM 30309(T)).

DOI： 10.1099/ijs.0.000356

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SPRINGER  

3-Methylindole, also referred to as skatole, is a pollutant of environmental concern due to its persistence, mobility and potential health impacts. Petroleum refining, intensive livestock production and application of biosolids to agricultural lands result in releases of 3-methylindole to the environment. Even so, little is known about the aerobic biodegradation of 3-methylindole and comprehensive biotransformation pathways have not been established. Using glycerol as feedstock, the soil bacterium Cupriavidus sp. strain KK10 biodegraded 100 mg/L of 3-methylindole in 24 h. Cometabolic 3-methylindole biodegradation was confirmed by the identification of biotransformation products through liquid chromatography electrospray ionization tandem mass spectrometry analyses. In all, 14 3-methylindole biotransformation products were identified which revealed that biotransformation occurred through different pathways that included carbocyclic aromatic ring-fission of 3-methylindole to single-ring pyrrole carboxylic acids. This work provides first comprehensive evidence for the aerobic biotransformation mechanisms of 3-methylindole by a soil bacterium and expands our understanding of the biodegradative capabilities of members of the genus Cupriavidus towards heteroaromatic pollutants.

DOI： 10.1007/s10532-015-9739-0

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1002/mbo3.283

PubMed

J-GLOBAL

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

添付ファイル： Kawsar et al (2015) Current Res Chem 7(2), 21-33.pdf

DOI： 10.3923/crc.2015.21.33

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

添付ファイル： Koide et al (2015) Ann. Marine Biol. Res. 2, 1005.pdf

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCI LTD  

Comprehensive analyses of the biotransformation of the N-heteroaromatic environmental pollutant, indole, by a newly isolated soil bacterium designated strain KK10, were conducted by liquid chromatography negative electrospray ionization tandem mass spectrometry (LC/ESI(-)-MS/MS). Numerous indole bioproducts were revealed and provided evidence for multiple pathways of indole biotransformation by this strain including unreported pathways. Oxidation of the N-heterocyclic ring of indole and ring cleavage through N-formylanthranilic acid and gentisic acid was characterized in addition to a carbocyclic ring-cleavage pathway which was documented for the first time. Three carbocyclic aromatic ring cleavage products of indole were proposed and the structure of 2,3-pyrrole-dicarboxylic acid was confirmed following synthesis and analyses of an authentic standard of this compound. Eight downstream ring-fission products were identified overall and these results confirmed the lower pathways of indole biotransformation. Indigo, indirubin and other indigoid compounds were produced by strain KK10 and confirmed N-heterocyclic-ring oxidations in the upper pathways. Nearly complete sequencing of the 16S rRNA gene of strain KK10 and phylogenetic analysis revealed that it was a new member of the genus Cupriavidus. Cupriavidus strains that biotransform N-heteroaromatic pollutants have not been described. These results expand both our understanding of the versatile catabolic capabilities of members of the genus Cupriavidus and provide evidence for alternative biotransformation mechanisms for indole in the environment by bacteria. (C) 2014 Elsevier Ltd. All rights reserved.

添付ファイル： Fukuoka et al (2015) Int Biodetrio Biodegra 97 13-24.pdf

DOI： 10.1016/j.ibiod.2014.11.007

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCI LTD  

Comprehensive analyses of the biotransformation of the N-heteroaromatic environmental pollutant, indole, by a newly isolated soil bacterium designated strain KK10, were conducted by liquid chromatography negative electrospray ionization tandem mass spectrometry (LC/ESI(-)-MS/MS). Numerous indole bioproducts were revealed and provided evidence for multiple pathways of indole biotransformation by this strain including unreported pathways. Oxidation of the N-heterocyclic ring of indole and ring cleavage through N-formylanthranilic acid and gentisic acid was characterized in addition to a carbocyclic ring-cleavage pathway which was documented for the first time. Three carbocyclic aromatic ring cleavage products of indole were proposed and the structure of 2,3-pyrrole-dicarboxylic acid was confirmed following synthesis and analyses of an authentic standard of this compound. Eight downstream ring-fission products were identified overall and these results confirmed the lower pathways of indole biotransformation. Indigo, indirubin and other indigoid compounds were produced by strain KK10 and confirmed N-heterocyclic-ring oxidations in the upper pathways. Nearly complete sequencing of the 16S rRNA gene of strain KK10 and phylogenetic analysis revealed that it was a new member of the genus Cupriavidus. Cupriavidus strains that biotransform N-heteroaromatic pollutants have not been described. These results expand both our understanding of the versatile catabolic capabilities of members of the genus Cupriavidus and provide evidence for alternative biotransformation mechanisms for indole in the environment by bacteria. (C) 2014 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.ibiod.2014.11.007

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Academic Journals Inc.  

Methyl α-D-glucopyranoside was easily prepared by the treatment of D-glucose with anhydrous methyl alcohol in presence of hydrogen chloride at freezing temperature in good yield. Then N-acetylsulfanilylation of methyl α-D-glucopyranoside has been carried out by the direct method and afforded the 6-O-N-acetylsulfanilyl derivative in an excellent yield. In order to obtain newer products, the 6-O-N-acetylsulfanilyl derivative was further transformed to a series of 2,3,4-tri-O-acyl derivatives containing a wide variety of functionalities in a single molecular framework. The chemical structures of the newly synthesized compounds were elucidated by Fourier Transform Infrared spectroscopy (FTIR), 1H-NMR (Proton nuclear magnetic resonance) spectroscopy elemental and physicochemical properties analysis. All the newly synthesized D-glucose derivatives were tested for their in vitro antibacterial activity against some human pathogenic bacterial strains. The study revealed that a good number of acylated products exhibited promising antibacterial activities. It is expected that the acylated derivatives of D-glucose may be considered as a potential source for developing new and better antibacterial agents against a number of pathogenic organism.

添付ファイル： Kawsar 2015 Int J Biol Chem 9(6) 269-282.pdf

DOI： 10.3923/ijbc.2015.269.282

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掲載種別：研究論文（学術雑誌）   出版者・発行元：The Society of Japanese Women Scientists  

DOI： 10.5939/sjws.15006

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記述言語：英語   出版者・発行元：SPRINGER  

DOI： 10.1007/s10695-014-9957-0

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記述言語：英語   出版者・発行元：OXFORD UNIV PRESS INC  

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SPRINGER  

Rhamnose-binding lectin (RBL) is one of the animal lectin categories which take part in the innate immune responses of fish. Osmerus lanceolatus lectin (OLL) from shishamo smelt eggs is an RBL composed of two tandem-repeated domains, both of which are considered to be a carbohydrate-recognition domain. SAL, catfish (Silurus asotus) egg RBL composed of three domains, binds to Burkitt's lymphoma Raji cells through globotriaosylceramide (Gb3) carbohydrate chain and to reduce cell size and growth by altering membrane composition without causing cell death. In this experiment, we tried to compare the binding effects of these two RBLs on Raji cells. Flow cytometric and fluorescence microscopic analyses revealed that OLL also directly bound to and shrunk Raji cells with ten times less reactivity than SAL but reduced cell growth with decreasing cell viability. Anti-Gb3 antibody completely blocked the binding of SAL to Raji cells but not that of OLL. In addition, the direct bindings of OLL and SAL to Raji cells were comparably inhibited by melibiose, but lactose was more effective inhibitor for the binding of OLL than that of SAL. These results suggest that OLL has slightly different cell-binding property compared with SAL and binds not only to Gb3 but also to the other carbohydrate receptor-bearing beta-galactoside chains. The quantitative RT-PCR analysis revealed that SAL induced the expression of TNF-alpha but not of IFN-gamma, IL-1 beta, and IL-10. Thus, SAL-induced cytostatic effect on Raji cells might be partially caused by TNF-alpha-mediated signaling pathway.

DOI： 10.1007/s10695-014-9948-1

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Rhamnose-binding lectin (RBL) is one of the animal lectin categories which take part in the innate immune responses of fish. Osmerus lanceolatus lectin (OLL) from shishamo smelt eggs is an RBL composed of two tandem-repeated domains, both of which are considered to be a carbohydrate-recognition domain. SAL, catfish (Silurus asotus) egg RBL composed of three domains, binds to Burkitt's lymphoma Raji cells through globotriaosylceramide (Gb3) carbohydrate chain and to reduce cell size and growth by altering membrane composition without causing cell death. In this experiment, we tried to compare the binding effects of these two RBLs on Raji cells. Flow cytometric and fluorescence microscopic analyses revealed that OLL also directly bound to and shrunk Raji cells with ten times less reactivity than SAL but reduced cell growth with decreasing cell viability. Anti-Gb3 antibody completely blocked the binding of SAL to Raji cells but not that of OLL. In addition, the direct bindings of OLL and SAL to Raji cells were comparably inhibited by melibiose, but lactose was more effective inhibitor for the binding of OLL than that of SAL. These results suggest that OLL has slightly different cell-binding property compared with SAL and binds not only to Gb3 but also to the other carbohydrate receptor-bearing β-galactoside chains. The quantitative RT-PCR analysis revealed that SAL induced the expression of TNF-α but not of IFN-γ, IL-1β, and IL-10. Thus, SAL-induced cytostatic effect on Raji cells might be partially caused by TNF-α-mediated signaling pathway.

添付ファイル： Hosono et al (2014) Fish Physiol Biochem 40 1559-1572.pdf

DOI： 10.1007/s10695-014-9948-1

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：MDPI AG  

A specific galactose-binding lectin was shown to inhibit the hemolytic effect of streptolysin O (SLO), an exotoxin produced by Streptococcus pyogenes. Commercially available lectins that recognize N-acetyllactosamine (ECA), T-antigen (PNA), and Tn-antigen (ABA) agglutinated rabbit erythrocytes, but had no effect on SLO-induced hemolysis. In contrast, SLO-induced hemolysis was inhibited by AKL, a lectin purified from sea hare (Aplysia kurodai) eggs that recognizes a-galactoside oligosaccharides. This inhibitory effect was blocked by the co-presence of D-galactose, which binds to AKL. A possible explanation for these findings is that cholesterol-enriched microdomains containing glycosphingolipids in the erythrocyte membrane become occupied by tightly stacked lectin molecules, blocking the interaction between cholesterol and SLO that would otherwise result in penetration of the membrane. Growth of S. pyogenes was inhibited by lectins from a marine invertebrate (AKL) and a mushroom (ABA), but was promoted by a plant lectin (ECA). Both these inhibitory and promoting effects were blocked by co-presence of galactose in the culture medium. Our findings demonstrate the importance of glycans and lectins in regulating mechanisms of toxicity, creation of pores in the target cell membrane, and bacterial growth.

添付ファイル： Hasan et al (2014) Molecules 19_13990-14003.pdf

DOI： 10.3390/molecules190913990

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：NATL INST SCIENCE COMMUNICATION-NISCAIR  

A new chitin-binding lectin was purified from a Bangladeshi cultivar `Deshi' of potato (Solanum tuberosum L.) through anion-exchange and affinity chromatographies using a chitin column. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) showed the molecular-mass of the lectin as 20,000 Daltons. This molecular mass was almost half of the molecular masses of chitin-binding lectins derived from other potatoes. The lectin showed both bactericidal and growth-inhibiting activities against Gram-positive (Listeria monocytogenes) and Gram-negative (Escherichia coli, Salmonella enteritidis and Shigella boydii) pathogenic bacteria. It also showed antifungal activity against Rhizopus spp., Penicillium spp. and Aspergillus niger. Biofilm produced by the bacterium Pseudomonas aeruginosa was dose-dependently reduced by 5-20% in 24 h after administration of the lectin, which was attributed to the glycan-binding property of the lectin having affinity to G1cNAc polymers. It was the first observation that any potato lectin prevented biofilm formation by P. aeruginosa and, therefore, could have possible applications in clinical microbiology and biomedical science.

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

文科省国費留学生Imtiaj Hasan氏(2013年3月YCU博士課程進学)の本邦初第一著者論文を出版できました。ポテトレクチンによるエールリッヒがん細胞死活性を報告

添付ファイル： Hasan et al (2014) African J Biotechnol 13(15) 1679-1685.pdf

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

研究指導博士院生Imtiaj Hasan筆頭論文第2報。キチン糖鎖結合性ポテトレクチンがバクテリアのバイオフィルム形成を阻害した新知見を報告

添付ファイル： Hasan et al (2014) Indian J Biochem Biophys 51(2) 142-148.pdf

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：WILEY  

A pathway for the biotransformation of the environmental pollutant and high-molecular weight polycyclic aromatic hydrocarbon (PAH) benzo[k]fluoranthene by a soil bacterium was constructed through analyses of results from liquid chromatography negative electrospray ionization tandem mass spectrometry (LC/ESI(-)-MS/MS). Exposure of Sphingobium sp. strain KK22 to benzo[k]fluoranthene resulted in transformation to four-, three- and two-aromatic ring products. The structurally similar four- and three-ring non-alternant PAHs fluoranthene and acenaphthylene were also biotransformed by strain KK22, and LC/ESI(-)-MS/MS analyses of these products confirmed the lower biotransformation pathway proposed for benzo[k]fluoranthene. In all, seven products from benzo[k]fluoranthene and seven products from fluoranthene were revealed and included previously unreported products from both PAHs. Benzo[k]fluoranthene biotransformation proceeded through ortho-cleavage of 8,9-dihydroxy-benzo[k]fluoranthene to 8-carboxyfluoranthenyl-9-propenic acid and 9-hydroxy-fluoranthene-8-carboxylic acid, and was followed by meta-cleavage to produce 3-(2-formylacenaphthylen-1-yl)-2-hydroxy-prop-2-enoic acid. The fluoranthene pathway converged with the benzo[k]fluoranthene pathway through detection of the three-ring product, 2-formylacenaphthylene-1-carboxylic acid. Production of key downstream metabolites, 1,8-naphthalic anhydride and 1-naphthoic acid from benzo[k]fluoranthene, fluoranthene and acenaphthylene biotransformations provided evidence for a common pathway by strain KK22 for all three PAHs through acenaphthoquinone. Quantitative analysis of benzo[k]fluoranthene biotransformation by strain KK22 confirmed biodegradation. This is the first pathway proposed for the biotransformation of benzo[k]fluoranthene by a bacterium.

DOI： 10.1111/1751-7915.12102

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Wiley  

ロバート・カナリー研究指導博士院生、前田亜鈴悠筆頭論文。環境保全微生物スフィンゴビウム属KK22株の多環芳香族毒物の分解代謝過程をLC/MSMSで同定。横浜市大連携大学院・海洋研究開発機構との共同研究

添付ファイル： 2013 Maeda Kanaly et al Microbial Biotechology.pdf

DOI： 10.1111/1751-7915.12102

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SPRINGER  

SBL/RC-RNase was originally isolated from frog (Rana catesbeiana) oocytes and purified as a novel sialic acid-binding lectin (SBL) that displayed strong anti-cancer activity. SBL was later shown to be identical to a ribonuclease (RC-RNase) from oocytes of the same species. The administration of SBL/RC-RNase induced apoptosis (with nuclear condensation and DNA fragmentation) in mouse leukemia P388 cells but did not kill umbilical vein endothelial or fibroblast cells derived from normal tissues. The cytotoxic activity of SBL/RC-RNase was inhibited by desialylation of P388 cells and/or the co-presence of free bovine submaxillary mucin. FACS analysis showed that SBL/RC-RNase was incorporated into cells after attachment to cholesterol-rich microdomains. Addition of the cholesterol remover methyl-beta-cyclodextrin reduced SBL/RC-RNase-induced apoptosis. Apoptosis occurred through the caspase-3 pathway following activation of caspase-8 by SBL/RC-RNase. A heat shock cognate protein (Hsc70) and a heat shock protein (Hsp70) (each 70 kDa) on the cell membrane were shown to bind to SBL/RC-RNase by mass spectrometric and flow cytometric analyses. Quercetin, an inhibitor of Hsc70 and Hsp70, significantly reduced SBL/RC-RNase-induced apoptosis. Taken together, our findings suggest that sialyl-glycoconjugates present in cholesterol-rich microdomains form complexes with Hsc70 or Hsp70 that act as triggers for SBL/RC-RNase to induce apoptosis through a pathway involving the activation of caspase-3 and caspase-8.

添付ファイル： Ogawa et al (2014) Glycoconj J 31(2) 171-184.pdf

DOI： 10.1007/s10719-013-9513-7

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Osterreichische Apotheker-Verlagsgesellschaft m.b.H.  

A regioselective acylation series of methyl α-D-glucopyranoside (1), methyl 3-O-benzoyl-4,6-O-benzylidene-α-D-mannopyranoside (1A), and methyl 4,6-O-benzylidene-2-O-(3,5-dinitrobenzoyl)-α-D-mannopyranoside (1B) has been carried out by the direct acylation method and afforded the 2,6-di-O-glucopyranoside and 2 or 3-O-mannopyranoside derivatives in an excellent yield. In order to obtain newer products, the 2,6-di-O-glucopyranoside derivative was further transformed to a series of 3,4-di-O-acyl derivatives containing a wide variety of functionalities in a single molecular framework. The structures of the newly synthesized compounds were elucidated on the basis of IR, 1H-NMR, 13C-NMR, 13C-DEPT spectral data, and elemental analysis. These synthesized derivatives were screened for in vitro antimicrobial activities against ten human pathogenic and five phytopathogenic microorganisms. A number of test compounds showed remarkable antimicrobial activity comparable to, and in some cases even higher than, the standard antibiotics employed. It was observed that methyl 3,4-di-O-(3-chlorobenzoyl)-2,6-di-O-hexanoyl-α-D-glucopyranoside (8) ex-hibited a varied range of MIC from 12.5 μg/disc to 25 μg/disc by the disk diffusion method and 1000 μg/mL to 1250 μg/mL by the broth macrodilution method. © Kawsar et al.

添付ファイル： Kawsar et al (2014) Sci Pharm 82 1-20.pdf

DOI： 10.3797/scipharm.1308-03

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Osmerus (Spirinchus) lanceolatus egg lectin (OLL) is a member of the rhamnose-binding lectin (RBL) family which is mainly found in aqueous beings. cDNA of OLL was cloned, and its genomic architecture was revealed. The deduced amino acid (aa) sequence indicated that OLL was composed of 213 aa including 95 aa of domain N and 97 aa of domain C. N and C showed 73 % sequence identity and contained both -ANYGR- and -DPC-KYL-peptide motifs which are conserved in most of the RBL carbohydrate recognition domains. The calculated molecular mass of mature OLL was 20,852, consistent with the result, and 20,677.716, from mass spectrometry. OLL was encoded by eight exons: exons 1 and 2 for a signal peptide; exons 3-5 and 6-8 for N- and C-domains, respectively. Surface plasmon resonance spectrometric analyses revealed that OLL showed comparable affinity for Galα- and β-linkages, whereas Silurus asotus lectin (SAL), a catfish RBL, bound preferentially to α-linkages of neoglycoproteins. The Kd values of OLL and SAL against globotriaosylceramide (Gb3) were 1.69 × 10⁻⁵ M for and 2.81 × 10⁻⁶ M, respectively. Thus, the carbohydrate recognition property of OLL is slightly different from that of SAL. On the other hand, frontal affinity chromatography revealed that both OLL and SAL interacted with only glycolipid-type oligosaccharides such as Gb3 trisaccharides, not with N-linked oligosaccharides. The domain composition of these RBLs and an analytical environment such as the "cluster effect" of a ligand might influence the binding between RBL and sugar chains.

添付ファイル： Hosono et al (2013) Fish Physiol Biochem 39 1619-1630.pdf

DOI： 10.1007/s10695-013-9814-6

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

JSPS海外特別研究員SMアベ・カウサル博士(現チッタゴン大学理学部化学科教授、受け入れ期間2009-11年)の帰国後初論文。有機合成した人工糖による顕著な抗菌効果を報告

添付ファイル： Kawsar et al (2013) Hacettepe J Biol Chem 41(3) 195-206.pdf

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：AMER SOC MICROBIOLOGY  

A soil bacterium, designated strain KK22, was isolated from a phenanthrene enrichment culture of a bacterial consortium that grew on diesel fuel, and it was found to biotransform the persistent environmental pollutant and high-molecular-weight polycyclic aromatic hydrocarbon (PAH) benz[a]anthracene. Nearly complete sequencing of the 16S rRNA gene of strain KK22 and phylogenetic analysis revealed that this organism is a new member of the genus Sphingobium. An 8-day time course study that consisted of whole-culture extractions followed by high-performance liquid chromatography (HPLC) analyses with fluorescence detection showed that 80 to 90% biodegradation of 2.5 mg liter(-1) benz[a]anthracene had occurred. Biodegradation assays where benz[a]anthracene was supplied in crystalline form (100 mg liter(-1)) confirmed biodegradation and showed that strain KK22 cells precultured on glucose were equally capable of benz[a]anthracene biotransformation when precultured on glucose plus phenanthrene. Analyses of organic extracts from benz[a]anthracene biodegradation by liquid chromatography negative electrospray ionization tandem mass spectrometry [LC/ESI(-)-MS/MS]revealed 10 products, including two o-hydroxypolyaromatic acids and two hydroxy-naphthoic acids. 1-Hydroxy-2- and 2-hydroxy-3-naphthoic acids were unambiguously identified, and this indicated that oxidation of the benz[a]anthracene molecule occurred via both the linear kata and angular kata ends of the molecule. Other two-and single-aromatic-ring metabolites were also documented, including 3-(2-carboxyvinyl)naphthalene-2-carboxylic acid and salicylic acid, and the proposed pathways for benz[a]anthracene biotransformation by a bacterium were extended.

添付ファイル： Kunihiro et al (2013) Appl Environ Microbiol 79 4410.pdf

DOI： 10.1128/AEM.01129-13

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記述言語：英語   掲載種別：論文集(書籍)内論文   出版者・発行元：John Wiley and Sons  

This chapter describes three aspects of marine invertebrate lectins based on glycomic studies of glycan-binding properties. The first section describes a lectin from the feather star, a close relative of the sea lily, as a molecular device to separate somatic and induced pluripotent stem cells (iPS cells). The second and third sections describe two lectins isolated from Mediterranean mussel and catfish eggs that have identical glycan-binding properties but completely different primary structures and different regulatory effects on Burkitt's lymphoma cells. © 2013 John Wiley &amp
Sons, Ltd.

添付ファイル： Ozeki et al (2013) Chapt 8 Marine Proteins & Peptides Wiley & Blackwell.pdf

DOI： 10.1002/9781118375082.ch8

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：BENTHAM SCIENCE PUBL LTD  

A highly cost-effective and easy-to-assemble cultivation system suitable for middle scale-size culturing of bacterial cells is described. In the culture, from a flat-shaped air-stone with large surface area, fine bubbles are generated with a low-cost air pump available in an aquarium fish shop, and cell-agitation and oxygen supply are efficiently conducted by fine bubbles simultaneously. Growth properties of the cells and their saturation density are comparable to those in a conventional culture system. The expression of recombinant protein was revealed to be similar to conventional methods. The system does not require any expensive machines or equipments. In addition, all equipments except plastic flat-shaped air-stone are reusable after sterilization. Due to the low cost, the ease to use and multiple cultivations at once, our system may enable to find better culture conditions, to scale-up with ease and to perform timesaving efficient protein production.

添付ファイル： Yasumitsu et al (2013) Protein Pep Lett 20 213-217.pdf

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：American Society for Microbiology  

Sphingobium sp. strain KK22 was isolated from a bacterial consortium that originated from cattle pasture soil from Texas. Strain KK22 grows on phenanthrene and has been shown to biotransform the high-molecular-weight (HMW) polycyclic aromatic hydrocarbon (PAH) benz[a]anthracene. The genome of strain KK22 was sequenced to investigate the genes involved in aromatic pollutant biotransformation.

添付ファイル： Maeda et al Kanaly e00911-13.full.pdf

DOI： 10.1128/genomeA.00911-13

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

A novel lectin structure was found for a 17-kDa alpha-D-galactose-binding lectin (termed "MytiLec") isolated from the Mediterranean mussel, Mytilus galloprovincialis. The complete primary structure of the lectin was determined by Edman degradation and mass spectrometric analysis. MytiLec was found to consist of 149 amino acids with a total molecular mass of 16,812.59 Da by Fourier transform-ion cyclotron resonance mass spectrometry, in good agreement with the calculated value of 16,823.22 Da. MytiLec had an N terminus of acetylthreonine and a primary structure that was highly novel in comparison with those of all known lectins in the structure database. The polypeptide structure consisted of three tandem-repeat domains of similar to 50 amino acids each having 45-52% homology with each other. Frontal affinity chromatography technology indicated that MytiLec bound specifically to globotriose (Gb3; Gal alpha 1-4Gal beta 1-4Glc), the epitope of globotriaosylceramide. MytiLec showed a dose-dependent cytotoxic effect on human Burkitt lymphoma Raji cells (which have high surface expression of Gb3) but had no such effect on erythroleukemia K562 cells (which do not express Gb3). The cytotoxic effect of MytiLec was specifically blocked by the co-presence of an alpha-galactoside. MytiLec treatment of Raji cells caused increased binding of anti-annexin V antibody and incorporation of propidium iodide, which are indicators of cell membrane inversion and perforation. MytiLec is the first reported lectin having a primary structure with the highly novel triple tandem-repeat domain and showing transduction of apoptotic signaling against Burkitt lymphoma cells by interaction with a glycosphingolipid-enriched microdomain containing Gb3.

添付ファイル： Fujii et al (2012) J Biol Chem 287 44772-44783.pdf

DOI： 10.1074/jbc.M112.418012

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：AMER CHEMICAL SOC  

Botrocetin is a heterodimer snake venom protein that induces von Willebrand factor (VWF)- and platelet glycoprotein Ib (GPIb)-dependent platelet agglutination in vitro. We have cloned cDNAs for a botrocetin-2 from a cDNA library of the venom gland of Bothrops jararaca having a high similarity with botrocetin subunits. Recombinant botrocetin-2, expressed in 293T cells, showed cofactor activity comparable to natural botrocetin. In a single subunit expression experiment, a dimer of the beta subunit was obtained, and it showed reduced, but apparent, platelet agglutination activity. Ala scanning mutagenesis showed that substitutions at Asp62, Asp70, Arg115, or Lys117 in the beta subunit reduced platelet agglutination activity. The 3D homology modeling of botrocetin-2 complexed with the VWF Al domain and GPIb alpha indicated that Asp62, Arg115, and Lys117 of the beta subunit are located near Arg218 and Asp222 of GPIb alpha, respectively, and that Asp beta 70 is in proximity to Gln1391 of the Al domain. Our results indicate that these charged amino acid residues in the beta subunit have a preferential role in the activity of botrocetin-2. Since it has been time-consuming and difficult to obtain homogeneous botrocetin from natural venom, recombinant botrocetin-2 has potential benefits for clinical and basic investigations into hemostasis and thrombosis as a standard reagent.

添付ファイル： Yamamoto et al Biochemistry(2012) 51 5329-5338.pdf

DOI： 10.1021/bi300442c

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：MDPI AG  

A divalent cation-independent lectin-HOL-18, with cytotoxic activity against leukemia cells, was purified from a demosponge, Halichondria okadai. HOL-18 is a 72 kDa tetrameric lectin that consists of four non-covalently bonded 18 kDa subunits. Hemagglutination activity of the lectin was strongly inhibited by chitotriose (GlcNAc beta 1-4GlcNAc beta 1-4GlcNAc), fetuin and mucins from porcine stomach and bovine submaxillary gland. Lectin activity was stable at pH 4-12 and temperatures lower than 60 degrees C. Frontal affinity chromatography with 16 types of pyridylaminated oligosaccharides indicated that the lectin had an affinity for N-linked complex-type and sphingolipid-type oligosaccharides with N-acetylated hexosamines and neuramic acid at the non-reducing termini. The lectin killed Jurkat leukemia T cells and K562 erythroleukemia cells in a dose- and carbohydrate-dependent manner.

添付ファイル： Matsumoto et al (2012) Toxins 4 323-338.pdf

DOI： 10.3390/toxins4050323

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記述言語：日本語  

J-GLOBAL

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記述言語：日本語   掲載種別：研究論文（学術雑誌）  

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SPRINGER  

A novel anticancer mechanism of catfish (Silurus asotus) egg lectin (SAL) was found to occur via the down-regulation of the membrane transopter protein, MRP1 (multidrug resistance associate protein-1) on Burkitt's lymphoma cells through Gb3(Gal alpha 1-4Gal beta 1-4Glc)-glycosphingolipid. Although SAL did not influence the viability of the cells directly, only 10 and 100 ng/mL of vincristine and etoposide, respectively induced anticancer effects when the lectin was applied in conjunction with these drugs. These phenomena were specifically inhibited by the co-presence of the alpha-galactoside, melibiose, which is a strong haptenic sugar of SAL that mimicks Gb3. The degree of expression regulation of the transporter proteins on the cells surface was investigated through the examination of the binding between SAL and Gb3-glycosphingolipid by immunological and molecular biological procedures. PCR data showed that MRP1 was more highly expressed when compared to another ATP-binding cassette family, multi-drug resistant protein and the expression levels of MRP1 on the cells were specifically dose- and time-dependently depleted by the addition of SAL. These results were also evaluated by immunological procedures using FACS and western-blotting. Small interfering RNA coding a part of MRP1 was transfected to Raji cells to knock down the protein, and cell death was increased by 10% when vincristine was administered at a concentration as low as 10 ng/mL compared to non-transfected cells. These results indicated that SAL possesses the potential to enhance the anticancer activites of low-concentrations of vincristine by the down-regulating the MRP1 gene expression to inhibit the multidrug resistance by binding to the target ligand Gb3-glycosphingolipid on Burkitt's lymphoma cells.

添付ファイル： Fujii et al (2012) Protein J 31,(1) 15-26.pdf

DOI： 10.1007/s10930-011-9369-2

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SPRINGER  

Galectin-1 from American bullfrog, RCG1, was isolated to high purity, and its growth inhibitory properties against human cells were examined. The results demonstrated that highly purified RCG1 induced large cell aggregates and revealed cell-type-specific growth inhibition. It significantly inhibited all human leukemia cell lines tested such as HL-60, U937, and K562 cells but did not inhibit human colon cancer cell line, Colo 201, or mouse mammary tumor cell line FM3A cells. Although most of the galectin-induced growth inhibitions are known to be apoptic, RCG1 induced growth arrest and neither apoptosis nor necrosis. RCG1-mediated growth inhibition was specifically suppressed by the corresponding sugar, lactose, but not by sucrose or even the structurally similar sugar, melibiose. Several studies have reported that galectin-mediated biological functions were modulated by charge modification. Since the high purity of RCG1 was demonstrated but a moderate degree of growth inhibition occurred, it is possible protein charge modification was examined by isoelectric focusing, and it was found to be highly heterogeneous in charge. RCG1 binding proteins in human cells were analyzed by lectin blotting using biotinylated RCG1, and lectin blotting revealed that in human cell extracts the specific proteins at molecular weight 37 and 50 kDa possessed the responsive features of RCG1 binding and lactose competition.

添付ファイル： Yasumitsu et al (2011) In Vitro Cell Dev Biol 47, 728-734.pdf

DOI： 10.1007/s11626-011-9462-z

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SPRINGER  

A divalent cation-independent 16 kDa d-galactose binding lectin (AKL-2) was isolated from eggs of sea hare, Aplysia kurodai. The lectin recognized d-galactose and d-galacturonic acid and had a 32 kDa dimer consisting of two disulfide-bonded 16 kDa subunits. Eighteen N-terminus amino acids were identified by Edman degradation, having unique primary structure. Lectin blotting analysis with horseradish peroxidase-conjugated lectins has shown that AKL-2 was a glycoprotein with complex type oligosaccharides with N-acetyl d-glucosamine and mannose at non-reducing terminal. Two protein bands with 38 and 36 kDa in the crude extract of sea hare eggs after purification of the lectin was isolated by AKL-2-conjugated Sepharose column and elution with 0.1 M lactose containing buffer. It suggested that the lectin binds with an endogenous ligand in the eggs. AKL-2 kept extreme stability on haemagglutination activity if it was treated at pH 3 and 70 A degrees C for 1 h. Glycan binding profile of AKL-2 by frontal affinity chromatography technology using 15 pyridylamine labeled oligosaccharides has been appeared that the lectin uniquely recognized globotriose (Gal alpha 1-4Gal beta 1-4Glc; Gb3) in addition to bi-antennary complex type N-linked oligosaccharides with N-acetyllactosamine. Surface plasmon resonance analysis of AKL-2 against a neo-glycoprotein, Gb3-human serum albumin showed the k (ass) and k (diss) values are 2.4 x 10(3) M(-1) s(-1) and 3.8 x 10(-3) s(-1), respectively. AKL-2 appeared cytotoxicity against both Burkitt&apos;s lymphoma Raji cell and erythroleukemia K562. The activity to Raji by the lectin was preferably cancelled by the co-presence of melibiose mimicing Gb3. On the other hand, K562 was cancelled effectively by lactose than melibiose. It elucidated that AKL-2 had cytotoxic ability mediated glycans structure to cultured cells.

添付ファイル： Kawsar et al (2011) Protein Journal 30 509-519.pdf

DOI： 10.1007/s10930-011-9356-7

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE INC  

A lectin - designated OXYL for the purposes of this study that strongly recognizes complex-type oligosaccharides of serum glycoproteins - was purified from a crinoid, the feather star Oxycomanthus japonicus, the most basal group among extant echinoderms. OXYL was purified through a combination of anion-exchange and affinity chromatography using Q-sepharose and fetuin-sepharose gel, respectively. Lectin was determined to be a 14-kDa polypeptide by sodium dodecyl sulphate-polyacrylamide gel electrophoresis under reducing conditions. However, 14-kDa and 28-kDa bands appeared in the same proportion under non-reducing conditions. Gel permeation chromatography showed a 54-kDa peak, suggesting that lectin consists of four 14-kDa subunits. Divalent cations were not indicated, and stable haemagglutination activity was demonstrated at pH 4-12 and temperatures below 60 degrees C. Surface plasmon resonance analysis of OXYL against fetuin showed k(ass), and k(diss) values of 1.4 x 10(-6) M(-1) s(-1) and 3.1 x 10(-3) s(-1), respectively, indicating that it has a strong binding affinity to the glycoprotein as lectin. Frontal affinity chromatography using 25 types of prydylamine-conjugated glycans indicated that OXYL specifically recognizes multi-antennary complex-type oligosaccharides containing type-2 N-acetyllactosamines (Gal beta 1-4GlcNAc) if alpha 2-3-linked sialic acid is linked at the non-reducing terminal. However, type-1 N-acetyllactosamine (Gal beta 1-3GlcNAc) chains and alpha 2-6-linked sialic acids were never recognized by OXYL This profiling study showed that OXYL essentially recognizes beta 1-4-linkage at C-1 position and free OH group at C-6 position of Gal in addition to the conservation of N-acetyl groups at C-2 position and free OH groups at C-3 position of GlcNAc in N-acetyllactosamine. This is the first report on glycornics on a lectin purified from an echinoderm belonging to the subphylum Pelmatozoa. (c) 2011 Elsevier Inc. All rights reserved.

添付ファイル： Matsumoto et al (2011) CBP 158 266-273.pdf

DOI： 10.1016/j.cbpb.2010.12.004

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE INC  

A divalent, cation-independent D-galactose-binding lectin was purified from coronate moon turban Turbo (Lunen) coreensis. This lectin recognizes D-galactose and is a 38-kDa dimeric protein consisting disulphide-bonded 22-kDa polypeptides under non-reducing and reducing conditions of sodium dodecyl sulphate-polyacrylamide gel electrophoresis, respectively. Haemagglutination activity was inhibited by D-galactose, N-acetyl D-galactosamine, melibiose, lactose, porcine stomach mucin, asialofetuin and bovine submaxillary mucin. The lectin has tolerance for pH 5-11 and temperature until 50 degrees C for 1 h. The lectin strongly aggregated Gram-negative bacteria, such as Vibrio parahaemolyticus and Salmonella 07, but weakly Gram-positive strain as Staphylococcus aureus and Bacillus subtilis. The glycan-binding profile of this lectin was evaluated using frontal affinity chromatography technology and the lectin appeared to recognize oligosaccharides such as lacto-series glycosphingolipids contained in blood type A and H substances in addition to complex-type N-linked glycoproteins. Partial primary structures of 139 amino acid residues of this lectin were determined from N-terminus polypeptides and 8 peptides derived by cleavage with lysyl-endopeptidase. The primary structure was slightly similar to other known sequences of lectin: however, a repeating motif has been included. (C) 2010 Elsevier Inc. All rights reserved.

添付ファイル： Fujii et al CBP 158 30-37 (2011).pdf

DOI： 10.1016/j.cbpb.2010.09.002

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掲載種別：研究論文（学術雑誌）  

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

添付ファイル： Kawsar et al (2011) Phytologia Balcanica 17, 347-354.pdf

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ACADEMIC PRESS INC ELSEVIER SCIENCE  

Coomassie Brilliant Blue (CBB) protein stains are inexpensive but detect proteins at only at microgram levels. Because of acetic acid and methanol, they cause skin irritation and reduce work motivation by malodor. Recent mass spectrometric (MS) analyses demonstrated that nanogram-sensitive colloidal CBB staining resulted in in vitro methylations of proteins. We propose a rapid, inexpensive, sensitive, odorless, less harsh, and in vitro methylation-free CBB stain. CGP uses three components: citric acid, CBB G-250, and polyvinylpyrrolidone. CGP detects proteins at 12 ng within 45 min, and because it is non-alcohol, in principle in vitro methylation would be eliminated. Indeed, MS analysis of CGP-stained bands confirmed a lack of methylation. (C) 2010 Elsevier Inc. All rights reserved.

添付ファイル： Yasumitsu et al (2010) Anal Biochem 406 86-88.pdf

DOI： 10.1016/j.ab.2010.06.035

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

添付ファイル： Kawsar et al (2010)Int J Biol Life Sci 6 31-37.pdf

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：WILEY-BLACKWELL  

SDS-PAGE and CBB staining are two of the most popular methods used for protein analysis. Although many reports that describe such staining methods have been published, these conventional protocols require several hours or days for staining and de-staining. In this study we describe a recently developed, fast and sensitive CBB staining method that utilizes the staining solution of RAMA that consists of the low-cost reagents: CBB R250, acetic acid, methanol and ammonium sulfate, and the destaining solution of water. Our method dose dependently detects 12 nanograms protein within 60 min and with a wide protein spectrum. Although the features of the dose-dependent relationship depend upon protein amounts and protein types, for most of the protein samples tested, a linear relationship was observed in the region from 12 to 330 ng. Moreover, through further washing, the detection sensitivity of protein is enhanced and reaches a maximum at 1.4 ng and then gradually decreases in the de-staining process. It has been shown recently through MS analyses that the sensitive colloidal CBB staining methods frequently result in artifactual methylations due to the strong acid and long contact during staining and the destaining processes. Such artifacts were reported to be reduced by the replacement of strong inorganic acid with acetic acid and because RAMA utilizes acetic acid and is in contact with the proteins for a short time during staining and destaining, it is expected that in vitro artifacts will be reduced. Finally, MS analyses of RAMA-stained protein bands were revealed not to have been methylated.

添付ファイル： Yasumitsu et al (2010) Electrophoresis 31 1913-1917.pdf

DOI： 10.1002/elps.200900524

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：BENTHAM SCIENCE PUBL LTD  

MMP-7 is the smallest metalloproteinase. Its unregulated activities and existence in serum are recently known to be tightly related with life-threatening disease such as cardiac disease and several cancers. The protein production is thought to be useful for its characterization and antibody generation. Although many attempts at bacterial expressions have been conducted, they were recovered as insoluble and inactive protein. In this study, after soluble expression, single-step purification and conversion to active protease, it was applied for the screening secretory metalloproteinase inhibitors in conditioned media of human cancer cells.

DOI： 10.2174/092986610791112648

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Zenodo  

Lectins have a good scope in current clinical
microbiology research. In the present study evaluated the
antimicrobial activities of a D-galactose binding lectin (PnL) was
purified from the annelid, Perinereis nuntia (polychaeta) by affinity
chromatography. The molecular mass of the lectin was determined to
be 32 kDa as a single polypeptide by SDS-PAGE under both reducing
and non-reducing conditions. The hemagglutinating activity of the
PnL showed against trypsinized and glutaraldehyde-fixed human
erythrocytes was specifically inhibited by D-Gal, GalNAc,
Galβ1-4Glc and Galα1-6Glc. PnL was evaluated for in vitro
antibacterial screening studies against 11 gram-positive and
gram-negative microorganisms. From the screening results, it was
revealed that PnL exhibited significant antibacterial activity against
gram-positive bacteria. Bacillus megaterium showed the highest
growth inhibition by the lectin (250 μg/disc). However, PnL did not
inhibit the growth of gram-negative bacteria such as Vibrio cholerae
and Pseudomonas sp. PnL was also examined for in vitro antifungal
activity against six fungal phytopathogens. PnL (100 μg/mL) inhibited
the mycelial growth of Alternaria alternata (24.4%). These results
indicate that future findings of lectin applications obtained from
annelids may be of importance to life sciences.

添付ファイル： Kawsar et al (2010)Int J Biol Life Sci 6 44-50.pdf

DOI： 10.5281/ZENODO.1057039

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掲載種別：研究論文（学術雑誌）  

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：INST BIOLOSKA ISTRAZIVANJA SINISA STANKOVIC  

The present investigation was undertaken in order to evaluate the cytotoxic effect of two D-galactose-binding lectins using the brine shrimp lethality bioassay technique. Both lectins were purified from the marine invertebrates, sea hare Aplysia kurodai eggs and polychaete Perineries nuntia by conventional affinity chromatography methods. The molecular mass of Aplysia kurodai egg lectin (AKL) was determined to be 32 kDa and 56 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis under reducing and non-reducing conditions, respectively. On the other hand, polychaete Perineries nuntia lectin (PnL) was determined to be 32 kDa in both reducing and non-reducing conditions. AKL and PnL showed strong agglutination activity against trypsinized and glutaraldehyde-fixed human and rabbit erythrocytes. AKL significantly affects the mortality rate of brine shrimp. Experimental results revealed that AKL was found to be more toxic (63.33% mortality) than PnL (33.33% mortality) and the mortality rate of brine shrimp nauplii was increased with the increase in concentration of lectins. These cytotoxic results indicate that future findings of lectin applications obtained front marine invertebrates may be of importance to clinical microbiology, and that they could have application as potent chemotherapeutic agents.

添付ファイル： Kawsar et al (2010) Arch Biol Sci Belgrade 62 1927-1038.pdf

DOI： 10.2298/ABS1004027K

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：MAIK NAUKA/INTERPERIODICA/SPRINGER  

A lectin was purified from Japanese sea hare Aplysia kurodai by lactosyl-agarose affinity chromatography. The molecular mass of the lectin was determined to be 56 and 32 kDa by SDS-PAGE under non-reducing and reducing conditions, respectively. It was found to agglutinate trypsinized and glutaraldehyde-fixed rabbit and human erythrocytes in the absence of divalent cations. The lectin exhibited stable thermo-tolerance as it retained hemagglutinating activity for 1 h even at 80A degrees C and showed stability at pH 10. By contrast, it was very sensitive at pH less than 5 and in the presence of the sulfhydryl-group preserving reagent, beta-mercaptoethanol. The hemagglutinating activity by the lectin was specifically inhibited by D-galactose, galacturonic acid, methyl-alpha- and methyl-beta-D-galactopyranoside, lactose, melibiose, and asialofetuin. The association rate constant (k (ass)) and dissociation rate constant (k (diss)) were determined for the lectin to be 4.3 center dot 10(5) M-1 center dot sec(-1) and 2.2 center dot 10(-3) sec(-1), respectively, using a surface plasmon resonance biosensor. The lectin moderately inhibited cell proliferation in the P388 cell line dose dependently. Interestingly, lectin-treated cells did not show a fragmented DNA ladder as is caused by apoptosis, suggesting that the cell proliferation inhibition was caused by another unknown mechanism.

添付ファイル： Kawsar et al (2009) Biochemistry(Moscow)74, 709-716.pdf

DOI： 10.1134/S0006297909070025

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：6  

産業総合研究所糖鎖医工学研究センターのレクチンデータベース構築のために提供したウシガエル卵ガレクチンの糖鎖結合プロファイルは、他のβガラクトシド結合性ガレクチンと異なり、キノコやカイメン由来のガレクチン同様に血液型A型やフォルスマン抗原などN-アセチルガラクトサミンンを特異的に認識し、細胞接着も行うことが見いだせられました

添付ファイル： Kawsar et al (2009) PPL 16,677-684.pdf

DOI： 10.2174/092986609788490104

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE INC  

A lectin recognizing D-galactose was purified from the pacific annelid Perinereis nuntia ver. vallata (Polychaeta) by affinity chromatography. Hemagglutinating activity, with a very low titer suggesting the presence of lectin appeared in the supernatant from the homogenization of body with Tris-buffered saline. However, dialyzed supernatant from the precipitate homogenized by galactose in the buffer revealed strong hemagglutinating activity against human erythrocytes. The crude supernatant was applied onto lactosylagarose column, and only the supernatant eluted from precipitate with galactose was obtained a galactose-binding lectin with 32 kDa polypeptide was obtained from the supernatant of the precipitate, extracted in presence of galactose. It suggests that the lectin tightly binds with glycoconjugate as endogenous ligand(s) in the tissue. Hemagglutinating activity against trypsinized and glutaraldehyde-fixed human erythrocytes was specifically inhibited by D-galactose, N-acetyl-D-galactosamine, lactose, melibiose, and asialofetuin. Glycan-binding profile of the lectin analyzed by frontal affinity chromatography shows that the lectin recognizes branched complex type N-linked oligosaccharides and both type 1 (Gal beta 1-3GlcNAc) and type 2 (Gal beta 1-4GlcNAc) lactosamine. The surface plasmon resonance study of the lectin against asialofetuin showed the k(ass) and k(diss) values are 5.14 x 10(4) M-1 s(-1) and 2.9 x 10(-3) s(-1), respectively. The partial primary structure of the lectin reveals 182 amino acids with novel sequence. (C) 2009 Elsevier Inc. All rights reserved.

添付ファイル： Kawsar et al (2009) CBP 152, 382-389.pdf

DOI： 10.1016/j.cbpb.2009.01.009

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

The bioactivity guided separation of the dichloromethane extract of the aerial parts of Macrotyloma uniflorum Linn, resulted in the isolation of methyl ester of hexadecanoic and ethyl ester of hexadecanoic acid mixture (I) and n-hexadecanoic acid (II). The structures of the isolated compounds were elucidated by spectroscopic analysis, including UV, IR, 1H-NMR, 13C-NMR and mass spectroscopy. In addition, the fractionated crude extract of 1-butanol exhibited the significant hemolytic activity by using mouse erythrocytes. © 2009 Academic Journals Inc.

DOI： 10.3923/ijbc.2009.42.48

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

To find novel carbohydrate-binding proteins (lectins) from marine invertebrates to understand the binding mechanism of the protein and to apply it for glycan-dependent diagnostics and/or glycoconjugates capture technology. A D-galactoside-binding lectin was purified from toot of bladder moon shell, Glossaulax didyma by lactosyl-agarose affinity chromatography. The crude supernatant by Tris-buffered saline had strong hemagglutination activity against trypsinized and glutaraldehyde-fixed human erythrocyte. However, the activity was not inhibited by any tested saccharides and chilete reagents. On the other hand, the dialyzed crude supernatant obtained from the precipitates with 100 mM lactose in Tris-buffered saline had also hemagglutination activity inhibited by β-galactoside and D-galactose. The lectin was purified with lactosyl-agarose affinity chromatography. The molecular mass of the lectin was determined to be 60 kDa by SDS-PAGE under reducing and non-reducing conditions and being a 60 kDa polypeptide monomer by gel permeation chromatography. The association-rate constant (kass) and dissociation-rate constant (kdiss) determined for the lectin against asialofetuin was determined as 5.4×104 M-1sec-1 and 7.2×10-3sec-1, respectively. Lectin-conjugated Sepharose gel captured asialofetuin and eluted it by lactose-containing buffer from the gel, indicating that the lectin could catch the asialoglycoprotein. It was concluded that a many amount of a D-galactoside-binding lectin which can catch asialoglycoprotein presents in foot of the bladder moon shell. © 2009 Asian Network for Scientific Information.

添付ファイル： Fujii et al (2009) J Biol Sci 9 (4) 319-325.pdf

DOI： 10.3923/jbs.2009.319.325

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DOI： 10.2174/092986609788490104

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE INC  

添付ファイル： Kawsar et al (2008) CBP 150 pp349-357.pdf

DOI： 10.1016/j.cbpb.2008.04.004

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE INC  

DOI： 10.1016/j.cbpb.2008.04.004

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Extracts of Macrotyloma uniflorum plants were examined as potential sources of phenolic compounds. Reversed phase high performance liquid chromatography (RP-HPLC) with UV detection was employed for the identification and quantification of the phenolic acids. Eight phenolic acids, namely, 3, 4-dihydroxy benzoic, p-hydroxy benzoic, vanillic, caffeic, p-coumeric, ferulic, syringic and sinapic acids were isolated from an ethanolic extract of Macrotyloma uniflorum. The most abundant phenolic acids were p-coumaric acid (8.95 mg 10-2 g of dry sample) and p-hydroxy benzoic acid (7.81 mg/100 g of dry sample). © 2008 Academic Journals Inc.

DOI： 10.3923/ajpp.2008.165.172

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

In the present research, attempt was taken to explore the antimicrobial potency of the crude extracts of the Macrotyloma uniflorum plant. The extractives of the plant were subjected to screening for inhibition of microbial growth by the disc diffusion method. The zones of inhibition demonstrated by the dichloromethane, ethyl acetate, 1-butanol and aqueous extracts ranged from 11-16, 10-24, 10-14 and 10-12 mm, respectively at a concentration of 500 μg disc-1. The ethyl acetate extract showed promising antibacterial activities against all the gram-positive and gram-negative bacteria whereas dichloromethane extract showed moderate activities and the 1-butanol and aqueous extracts did not show any significant antimicrobial activities. In addition, the antifungal activities of all the extractives were tested, using the food poisoning technique. Only dichloromethane extract has been proved to be active against all fungi tested with a higher inhibition activity than standard nystatin. The overall results provide promising baseline information for the potential use of the crude extracts from M. uniflorum in the treatment of bacterial and fungal infections. © 2008 Asian Network for Scientific Information.

DOI： 10.3923/jbs.2008.1051.1056

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE INC  

Galactoside-binding lectin was purified from the snake venom of Crotalus ruber by affinity chromatography on a lactose-agarose column, and the complete amino acid sequence was determined. The C. ruber venom lectin (CRL) showed a single band of 28 kDa by SDS-polyacrylamide electrophoresis under non-reducing conditions, but it showed a single band of 15 kDa under reducing conditions, indicating that CRL is a disulfide-linked homodimer of 15 kDa subunit. CRL specifically recognized beta-galactosides such as thiodigalactoside followed by Nacetylgalactosamine when examined with their inhibitory effects on CPL-induced hemagglutination. A CRL subunit was composed of 135 residues containing nine Cys residues and showed a high similarity to other C-type galactoside-binding lectins from snake venoms. C. atrox lectin (CAL) showed almost the same sequence except for eight amino acid residues. Neither CRL nor CAL induced platelet aggregation by itself or inhibited platelet aggregation mediated by von Willebrand factor or fibrinogen with agonists. CRL showed a similar oligomeric form and the sugar specificity as CAL, but it showed different divalent cation sensitivity such as Mn2+ and Ni2+. Homology modeling suggested that the amino acid substitution found in CRL does not affect sugar recognition of the lectin but might alter the conformation and influence the sugar binding pocket induced by the metal-ion binding. (c) 2006 Elsevier Inc. All rights reserved.

添付ファイル： Hamako et al (2007) Comp Biochem Physiol partB146 299-306.pdf

DOI： 10.1016/j.cbpb.2006.11.022

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE INC  

Galactoside-binding lectin was purified from the snake venom of Crotalus ruber by affinity chromatography on a lactose-agarose column, and the complete amino acid sequence was determined. The C. ruber venom lectin (CRL) showed a single band of 28 kDa by SDS-polyacrylamide electrophoresis under non-reducing conditions, but it showed a single band of 15 kDa under reducing conditions, indicating that CRL is a disulfide-linked homodimer of 15 kDa subunit. CRL specifically recognized beta-galactosides such as thiodigalactoside followed by Nacetylgalactosamine when examined with their inhibitory effects on CPL-induced hemagglutination. A CRL subunit was composed of 135 residues containing nine Cys residues and showed a high similarity to other C-type galactoside-binding lectins from snake venoms. C. atrox lectin (CAL) showed almost the same sequence except for eight amino acid residues. Neither CRL nor CAL induced platelet aggregation by itself or inhibited platelet aggregation mediated by von Willebrand factor or fibrinogen with agonists. CRL showed a similar oligomeric form and the sugar specificity as CAL, but it showed different divalent cation sensitivity such as Mn2+ and Ni2+. Homology modeling suggested that the amino acid substitution found in CRL does not affect sugar recognition of the lectin but might alter the conformation and influence the sugar binding pocket induced by the metal-ion binding. (c) 2006 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.cbpb.2006.11.022

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Axonal transport of endopeptidase 24.15 (EP24.15), a putative neuropeptide degrading-enzyme, was examined in the proximal, middle, and distal segments of rat sciatic nerves using a double ligation technique. At 48h after ligation, a significant amount of the axonal transport of EP24.15 activity was found in the proximal segment, while axonal transport of deamidase activity, a lysosomal enzyme, increased in both proximal and distal segments. Western blot analysis of EP24.15 showed that EP24.15 immunoreactivity in the proximal segment was 1.8-fold higher than that in the middle segment. The immunohistochemical analysis of the segments also showed an increase in the immunoreactive EP24.15 in the proximal segment in comparison with that in the middle segment. In the distal segment, no axonal transport of EP24.15 was found in all methods examined, indicating that EP24.15 is mainly transported by an anterograde axonal flow. These observations suggest that EP24.15 may be involved in the metabolism of neuropeptides in nerve terminals or synaptic clefts. © 2003 Elsevier Science Ltd. All rights reserved.

DOI： 10.1016/S0197-0186(02)00092-X

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE BV  

Binding specificities of ABO blood group-recognizing lectins toward blood group antigens on neoglycoproteins, glycoproteins and complex-type oligosaccharides were studied by lectin-blotting analysis, enzyme linked immunosorbent assay and lectin-conjugated agarose column chromatography. Human serum albumin conjugated with A- and B-trisaccharides was clearly recognized by Helix pomatia (WA), Phaseolus lunatus, Dolichos biflorus agglutinins, and Griffonia simplicifolia I agglutinin B-4, respectively. Almost the same results were obtained for human group A and B ovarian cyst and A-active hog gastric mucins, but Glycine max agglutinin only reacted to the group A hog mucin. When human plasma von Willebrand factor (VWF), having Asn-linked blood group antigens, was tested, HPA was highly sensitive to blood group A antigen on the VWF. Ulex europaeus agglutinin I (UEA-I) preferentially bound to the vWF from blood group O plasma. Within the GalNAc-recognizing lectins examined, a biantennary complex-type oligosaccharide having the blood group A structure retarded on an HPA-agarose column, and the affinity was diminished after digestion with alpha -N-acetylgalactosaminidase. This product bound to UEA-I agarose column. These results indicate that HPA and UEA-I are most sensitive for detection of glycoproteins possessing small amounts of blood group A and H antigens and also useful for fractionation of complex-type oligosaccharides with blood group A and H antigens, respectively. (C) 2001 Elsevier Science B.V. All rights reserved.

添付ファイル： Matsui et al (2001) Biochem Biophys Acta 1525 50-57.pdf

DOI： 10.1016/S0304-4165(00)00170-7

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：F K SCHATTAUER VERLAG GMBH  

A von Willebrand factor (vWF)-binding and -cleaving metalloproteinase, termed "kaouthiagin", was purified from the venom of cobra snake Naja kaouthia. Kaouthiagin is a monomer with a molecular mass of about 46 kDa and 51 kDa under non-reducing and reducing conditions, respectively, and the N-terminal amino acid sequence is homologous to high molecular mass snake venom metalloproteinases. Kaouthiagin bound to VWF in a divalent ion-independent manner, but the reduced kaouthiagin failed to interact with VWF, suggesting that the protein conformation maintained by intrachain-disulfide linkages of the molecule is essential for the binding to VWF. Neither botrocetin nor bitiscetin, vWF-binding modulators from another snake venom, interfered with the binding between kaouthiagin and vWF, but a monoclonal antibody VW92-3 specific to the N-terminal region of VWF (residues 1-910) inhibited the binding. Without affecting platelet GPIb/IX and GPIIb/IIIa, kaouthiagin specifically cleaved vWF between residues Pro-708 and Asp-709 in a divalent ion-dependent manner to diminish the multimeric structure of vWF in plasma, resulting in the loss of ristocetin-induced platelet aggregability and the collagen-binding activity of VWF. These results indicate that kaouthiagin is a unique metalloproteinase which specifically binds to and cleaves VWF at a specific site and that it will be a useful tool for functional dissection of VWF.

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Wiley-Blackwell  

添付ファイル： Ozeki (1998) IUBMB Life 45 989-995.pdf

DOI： 10.1002/iub.7510450516

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：OXFORD UNIV PRESS  

Galectins are a family of lectins that recognize beta-D-galactosides independently of calcium ions, and are widely distributed in animals, To characterize a galectin previously purified from oocytes of Rana catesbeiana (American bullfrog), we studied its distribution and localization in several tissues from this frog, Hemagglutination assay and western blotting showed that this lectin is present in many tissues including the liver, skin, kidney, skeletal muscle, and sciatic nerve, but is particularly concentrated in the ovary, Light microscopic immunohistochemistry showed that this lectin is localized in such places as cell-cell junctions, basement membranes, extracellular matrix, or secretory substances in several organs, indicating that this galectin is mainly distributed extracellularly, However, in the ovary, light microscopy showed that this lectin is present in or associated with the yolk platelet, Electron microscopy further revealed that it is localized in the periphery of the yolk platelet (the yolk plasm), but not in the cortical granule, These results indicate that Rana oocytes contain abundant galectin in their yolk platelets in contrast to Xenopus laevis oocytes, which have been found not to contain galectins but other classes of lectins in their yolk platelets and cortical granules.

添付ファイル： Uchiyama et al (1997) Glycobiology 7 1159-1165.pdf

DOI： 10.1093/glycob/7.8.1159

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記述言語：英語   出版者・発行元：W B SAUNDERS CO  

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD  

Lectins recognizing D-galactosides were purified from the body wall of an echiuroid; Urechis unicinctus and two annelids; Neanthes japonica and Marphysa sanguinea, with single step lactosyl-agarose affinity column chromatography. SDS-PAGE under reduced and non-reduced conditions showed that U, unicinctus lectin had a major (36 kDa) and two minor (40 and 14 kDa) proteins, and that N. japonica lectin and M. sanguinea lectin had single 33 and 35 kDa proteins, respectively. Lectins were solubilized in the presence of lactose from tissues, and all polypeptides were shown to have sugar binding activity. The antisera raised against U. unicinctus lectin and N. japonica Lectin crossreacted with each other but did nor crossreact with bull frog (Rana catesbeiana) egg galectin-1 or a D-galactoside specific lectin purified from sea urchin (Anthocidaris crassispina) eggs. These echiuroid and annelid lectins are immunologically similar, but distinct from members of the vertebrate galectin family. (C) 1997 Elsevier Science Inc.

添付ファイル： Ozeki et al (1997) CBP 118B 1-6.pdf

DOI： 10.1016/S0305-0491(97)00014-X

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ACADEMIC PRESS AUST  

A beta-D-galactoside binding lectin (TPL) was newly purified from the body walls of the cuttlefish, Todarodes pacificus with lactosyl-agarose affinity column chromatography and reversed-phase HPLC. Many of the biological properties of the lectin were similar to those of the representative animal beta-D-galactoside binding lectin, ''galectin''. However, with a molecular mass of 63 kDa under reducing and non-reducing conditions on SDS-PAGE, TPL is larger than any previously reported galectins. The native molecular mass of TPL was estimated to be about 60 kDa by gel filtration, suggesting that it exists as a monomer and that a single polypeptide containing at least two sugar binding sites. The antisera raised against TPL did not crossreact with bull frog egg galectin-1, suggesting that TPL is immunologically distinct from the lower vertebrate galectin.

添付ファイル： Ozeki Y (1997) IUBMB Life 41(3) 633-640.pdf

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

添付ファイル： Matsui et al (1997) J Biochem121 376-381.pdf

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：JAPANESE BIOCHEMICAL SOC  

The binding of human von Willebrand factor (vWF) to a variety of extracellular matrix components immobilized on plates and the binding of vWF to platelet glycoprotein It, (GPIb) after interacting with these matrix components mere examined by means of an enzyme-linked immunosorbent assay. vWF preferably bound to type III collagen, whereas it did not significantly bind to type I, IV, V, or VI collagen, fibronectin, laminin, elastin, or proteoglycans. Soluble type III collagen did not bind to vWF coated on plates and showed a little effect on the vWF binding to the immobilized collagen, suggesting that solid-phase collagen is important for the interaction with vWF. When glycocalicin, the N-terminal carbohydrate-rich extracellular domain of GPIb alpha exhibiting the vWF-binding activity, was added to vWF bound to collagen type III, no significant binding of glycocalicin was observed, but it bound to vWF in the presence of botrocetin, a vWF modulator protein isolated from Bothrops jararaca snake venom. These results indicate that vWP immobilized on collagen can interact with GPIb but that the binding of vWF to the collagen matrix alone is insufficient for modulating vWF so that it interacts with GPIb under static conditions. Another unknown physiological modulator functionally mimicking botrocetin or high-shear stress may be involved in the platelet adhesion to extracellular mat-fix in the early stage of hemostasis.

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ROCKEFELLER UNIV PRESS  

The structure of the carbohydrate of the 40-kD major outer membrane component of Chlamydia trachomatis and its role in defining infectivity of the organism were investigated. The oligosaccharides were released from the glycoprotein by N-glycanase digestion, coupled to a 2-aminopyridyl residue, and subjected to two-dimensional sugar mapping technique, The major fractions consisted of ''high-mannose type'' oligosaccharides containing 8-9 mannose residues, Bi- and tri-antennary ''complex type'' oligosaccharides having terminal galactose were detected as minor components. These oligosaccharides were N-linked and contained no sialic acid. This structural profile is consistent with our previous characterization based on lectin-binding and glycosidase digestion. Functional specificity of identified chlamydial oligosaccharides was analyzed using glycopeptides fractionated from ovalbumin and structurally defined oligosaccharides from other sources. The glycopeptide fraction having high-mannose type oligosaccharide, as compared to those having complex or hybrid-type, showed a stronger inhibitory effect on attachment and infectivity of chlamydial organisms to HeLa cells, Among high-mannose type oligosaccharides, the strongest inhibition was observed with mannose 8 as compared with mannose 6, 7, or 9. These results indicate that a specific high-mannose type oligosaccharide linked to the major outer membrane protein of C. trachomatis mediates attachment and infectivity of the organism to HeLa cells.

添付ファイル： Kou et al J Clin Invest 98(12)2813-2813.pdf

DOI： 10.1172/JCI119109

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS  

We have screened 20 snake venoms and purified a novel snake venom protein, named bitiscetin, from Bitis arietans venom that specifically binds to human von Willebrand factor (vWF) and induces platelet agglutination. Bitiscetin showed a heterodimeric structure composed of disulfide-linked alpha (16kDa) and beta (13kDa) subunits on SDS-PAGE and showed a basic nature with pI value of 9.1, in contrast to botrocetin (pI 4.6), a vWF modulator isolated from another snake (Bothrops jararaca) venom. Bitiscetin-induced platelet agglutination was dependent on vWF and platelet membrane glycoprotein (GP) Ib, bur not on Ca2+ and GPIIb/IIIa. vWF bound to bitiscetin but not to botrocetin electroblotted to a PVDF membrane after SDS-PAGE and this binding was diminished after reduction of disulfide bonds of bitiscetin. Bitiscetin did not cross-react to anti-botrocetin monoclonal antibodies. These results suggest that bitiscetin directly interacts with vWF and requires the protein conformation for its interaction as well as botrocetin, but its interaction manner with VWF appears to be different from that of botrocetin. (C) 1996 Academic Press, Inc.

DOI： 10.1006/bbrc.1996.1345

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掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/0305-0491(95)00121-2

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD  

The binding of IgM from a rheumatoid factor (RF-IgM) to IgG from 12 animal species was analyzed by an ELISA system. The RF-IgM bound various animal IgG with dissimilar affinities. The binding of RF-IgM to animal IgG was inhibited by addition of protein A, which binds some animal IgG by recognizing the junctional site on CH2-CH3 domains in the Fc region. As previously reported, no significant correlation was observed between the binding of RF-IgM to IgG and the content of galactose-free oligosaccharides, which is increased in IgG of rheumatoid arthritis patients or autoimmune mice. We suggest that the crucial epitope of IgG for RF-IgM binding is not the oligosaccharide structure generated specifically in IgG of antoimmune diseases but that RF-IgM may recognize a certain protein conformation of a region in IgG near the binding site of protein A.

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

添付ファイル： Hamako et al (1993) Comp Biochem Physiol 106B 949-954.pdf

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

A 14K β-galactoside-binding lecttn (galectin-1) is present in many animal tissues. In a search for endogenous ligands, we surveyed galectin-1-binding proteins in human placenta. Extract of human placenta with 2 M urea was applied to a Sepharose 4B column conjugated with galectin-1 purified from frog (Rana catesbeiana) eggs. Two major proteins eluted with 100 mM lactose from the column-bound fraction showed apparent molecular masses of 220 and 180 kDa on SDS-PAGE under reducing conditions. Western blotting analysis using monoclonal antibodies indicated that these proteins were fibronectin and laminin, respectively. Most placenta] and amniotic fibronectins bound strongly to the column, whereas almost all plasma fibronectin passed through the column. The galectin-1, fibronectin and laminin were immunohistochemically shown to be co-localized in the extracellular matrix of placental tissue. In a cell attachment assay, rhabdosarcoma cells adhered to a plate coated with placental fibronectin, even in the presence of GRGDS peptide, if galectin-1 were also present This adhesive effect of galectin-1 was inhibited by lactose. These results indicate that tissue fibronectin, as well as laminin, serve as endogenous ligands for galectin-1, suggesting that galectin-1 may play a role in assembly of the extracellular matrix, or in the control of cell adhesion based on lectin-extracellular matrix interaction. © 1995 Oxford University Press.

添付ファイル： Ozeki et al (1995) Glycobiology 5, 255-261.pdf

DOI： 10.1093/glycob/5.2.255

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS  

The spatial and temporal expression of a sea urchin (Anthocidaris crassispina) egg lectin (SUEL) during early embryogenesis was studied using antiserum raised against SUEL. Western blotting analysis revealed the presence of SUEL in all stages so far examined, from unfertilized eggs to gastrula stage embryos. Immunofluorescence and immunoelectron microscopic observation showed that SUEL was stored in small electron-dense granules which migrated to the cortex within 10 min after fertilization. SUEL was localized in the cortical cytoplasm of the blastomere during cleavage stages and subsequently migrated to the outer surface of the embryo, including the invaginated portion of the gastrula. Immunoelectron microscopic study indicated that SUEL was deposited in the hyaline layer at least at the mid gastrula stage. Migration of SUEL to the cortex was significantly reduced by treatment with cytochalasin B, suggesting that actin filaments play an important role in this translocation. Exogenously added SUEL was adsorbed at the surface of unfertilized eggs and hatched embryos, but not to embryos with fertilization membrane. Lactose inhibited this adsorption, suggesting the presence of an endogenous glycoligand(s) specific for SUEL on the surface of unfertilized eggs and in the hyaline layer. We conclude that SUEL is secreted at a certain stage of embryogenesis and specifically adsorbed to the hyaline layer. Temporal changes in extraembryonic matrices caused by SUEL seem to play an important role in developmental morphogenesis. (C) 1995 Academic Press, Inc.

添付ファイル： Ozeki et al (1995) Exp Cell Res 216, 318-324.pdf

DOI： 10.1006/excr.1995.1040

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS  

添付ファイル： Ozeki et al (1994) Arch Biochem Biophys 308(1) 306-310.pdf

DOI： 10.1006/abbi.1994.1043

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Oxford University Press  

Two structurally distinct lectins were purified from the coelomic plasma of holothurian, Stichopus japonicus, by affinity chromatography on a porcine stomach mucin-conjugated agarose column, gel filtration on a Superose 6 column, and ion-exchange chromatography on a HiTrap Q-FPLC. The two lectins showed apparent molecular masses of about 400 kDa (SPL-1) and 60 kDa (SPL-2) on gel filtration, but about 17 kDa on SDS-PAGE under reducing conditions. Both lectins showed hemagglutination activity toward rabbit eryth-rocytes in the presence of Ca2+ ions. The N-terminal amino acid sequences were highly homologous to but distinct from those of a Ca2+-dependent (C-type) lectin named SJL-I purified from the same species. In addition to porcine stomach mucin, the hemagglutination activity of SPL-1 was strongly inhibited by uronic acids such as galacturonic acid, and glucuronic acid, while the activity of SPL-2 was inhibited by GalNAc and galactosides. Both lectins were adsorbed on clotted coelomocytes in the presence of Ca2- but not in the presence of inhibitory sugars or EGTA, suggesting the presence of an endogenous carbohydrate ligand(s) for plasma C-type lectins in the clot. However, coelomocyte clotting occurred normally even in the presence of inhibitory sugars, but was strongly inhibited by synthetic GRGDSP peptide or EGTA, suggesting the participation of integrin but not the lectin-carbohydrate interaction in the clotting events. © 1994 BY THE JOURNAL OF BIOCHEMISTRY.

添付ファイル： Matsui et al (1994) J Biochem116 1127-1133.pdf

DOI： 10.1093/oxfordjournals.jbchem.a124638

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD  

1. Asparagine-linked sugar chains released by hydrazinolysis from IgGs of porcine, equine, bovine, goat, ovine, canine, rabbit, guinea-pig and rat were comparatively analyzed by microsequencing and lectin affinity chromatography.
2. Sugar chains of all IgGs basically consisted of biantennary complex-type oligosaccharides containing 0-2 sialic acid residue(s). More than 70% of the oligosaccharides were neutral, except for guinea-pig IgG, and fucosylated trimannosyl core structures were dominant except for rabbit IgG. Bisecting N-acetylglucosamine residue was absent in porcine and equine IgGs.
3. A large quantity of galactose-less oligosaccharides were present in IgGs of porcine, equine, canine and rat.

DOI： 10.1016/0305-0491(93)90056-B

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Elsevier {BV}  

添付ファイル： Hamako et al (1993) Comp Biochem Physiol 106B 949-954.pdf

DOI： 10.1016/0305-0491(93)90056-b

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：NATL ACAD SCIENCES  

The complete amino acid sequence and location of the disulfide bonds of two-chain botrocetin, which promotes platelet agglutination in the presence of von Willebrand factor, from venom of the snake Bothrops jararaca are presented. Sequences of the alpha and beta subunits were determined by analysis of peptides generated by digestion of the S-pyridyl-ethylated protein with Achromobacter protease I or alpha-chymotrypsin and by chemical cleavage with cyanogen bromide or 2-(2'-nitrophenylsulfenyl)-3-methyl-3-bromoindolenine. Two-chain botrocetin is a heterodimer composed of the alpha subunit (consisting of 133 amino acid residues) and the beta subunit (consisting of 125 amino acid residues) held together by a disulfide bond. Seven disulfide bonds link half-cystine residues 2 to 13, 30 to 128, and 103 to 120 of the alpha subunit; 2 to 13, 30 to 121, and 98 to 113 of the beta subunit; and 80 of the alpha subunit to 75 of the beta subunit. In terms of amino acid sequence and disulfide bond location, two-chain botrocetin is homologous to echinoidin (a sea urchin lectin) and other C-type (Ca2+-dependent) lectins.

添付ファイル： Usami et al (1993) PNAS 90 928-932.pdf

DOI： 10.1073/pnas.90.3.928

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE BV  

Cell adhesive activity of two animal lectins, frog (Rana catesbeiana) S-type 14K lectin and echinoidin (a C-type lectin from sea urchin plasma), was studied with human rhabdomyosarcoma (RD) cells. RD cells attached to and spread on plastic plates coated with each lectin. Cell adhesion by the frog lectin was completely inhibited by the addition of lactose or asialofetuin glycopeptide. Echinoidin-induced cell adhesion was only inhibited by peptide GRGDS. Since echinoidin is known to contain an RGD-sequence, our results clearly indicate that this sequence is active as the cell adhesive signal. These results suggest that some of the animal lectins may function as a cell adhesive molecule rather than using the carbohydrate-recognition mechanism.

DOI： 10.1016/0014-5793(91)81055-D

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

添付ファイル： Ozeki et al (1991) BBRC 178 404-413.pdf

DOI： 10.1016/0006-291X(91)91828-Z

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

The complete amino acid sequence of a 11.5-kDa subunit of D-galactoside binding lectin purified from sea urchin (Anthocidaris crassispina) eggs is presented. The 105-residue sequence of the subunit was determined by analysis of the intact S-carbamoylmethylated protein and peptides generated by digestion with Achromobacter protease I or Staphylococcus aureus V8 protease. The lectin exists as a disulfide-linked homodimer of two subunits
the dimeric form is essential for hemagglutination activity. However, the monomeric form obtained by partial reduction retains the carbohydrate binding capacity. Neither Ca2+ nor SH reagent is essential for hemagglutination or carbohydrate binding. The sequence has no similarity to that of any known protein and apparently represents a new type of galactoside binding lectin. © 1991, American Chemical Society. All rights reserved.

添付ファイル： Ozeki et al (1991) Biochemistry 30 2391-2394.pdf

DOI： 10.1021/bi00223a014

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：JAPANESE SOC BREEDING  

添付ファイル： Hattori et al (1990) J. J. Breed 40, 295-301.pdf

DOI： 10.1270/jsbbs1951.40.295

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総ページ数：540   担当ページ：407-418   記述言語：英語   著書種別：学術書

ASIN

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総ページ数：744   担当ページ：407-418   記述言語：英語   著書種別：学術書

ASIN

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総ページ数：816   担当ページ：167-184   記述言語：英語   著書種別：学術書

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記述言語：英語   会議種別：口頭発表（招待・特別）  

開催地：インド共和国 マハーラーシュトラ州 ムンバイ大学  

添付ファイル： Final Program 102nd ISC.pdf

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記述言語：日本語   会議種別：公開講演，セミナー，チュートリアル，講習，講義等  

添付ファイル： 2012年3月6日ライフイノベーションのための糖鎖関連シーズ神奈川県受託事業講演.pdf

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記述言語：日本語   会議種別：公開講演，セミナー，チュートリアル，講習，講義等  

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記述言語：日本語   会議種別：シンポジウム・ワークショップ パネル（指名）  

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記述言語：英語   会議種別：口頭発表（一般）  

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記述言語：英語   会議種別：ポスター発表  

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記述言語：英語   会議種別：ポスター発表  

開催地：Honolulu, Hawaii  

添付ファイル： Hasan et al (2014) Glycobiology 24 1053-1231 Conference abstract.pdf

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記述言語：日本語   会議種別：口頭発表（招待・特別）  

開催地：長崎県佐世保市  

添付ファイル： 2014年9月11日長崎国際大学要旨大関泰裕.pdf

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記述言語：英語   会議種別：口頭発表（招待・特別）  

開催地：インド共和国、ベンガルール  

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添付ファイル： 2019-06-08_.jpg

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記述言語：英語   会議種別：口頭発表（一般）  

開催地：Baltimore, Maryland, USA  

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会議種別：口頭発表（招待・特別）  

開催地：秋葉原コンベンションホール５階カンファレンスフロア/５B会議室  

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会議種別：シンポジウム・ワークショップ パネル（指名）  

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添付ファイル： 2018_June_09 CGM.jpg

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記述言語：日本語   会議種別：口頭発表（一般）  

添付ファイル： 2017_June_10 CGM.jpg

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会議種別：公開講演，セミナー，チュートリアル，講習，講義等  

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記述言語：英語   会議種別：口頭発表（一般）  

開催地：中国、青島市  

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発表場所：Essentials of Glycobiology  

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その他リンク： https://www.ncbi.nlm.nih.gov/books/NBK579951/

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発表場所：横浜市庁舎アトリウム  

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Ch 31 R-type Lectins
Richard D Cummings, Ronald L Schnaar, & Yasuhiro Ozeki

DOI： 10.1101/9781621824213

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発表場所：横浜市立大学  

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添付ファイル： 第23回比較グライコーム研究会2021ポスターfinal.pdf

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Collaboration of lectin researches with crystallographers

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その他リンク： https://www2.chubu.ac.jp/digibook/wpwt/216/html5.html#page=15

発表場所：横浜市立大学  

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添付ファイル： onlineシンポ2020年8月21日比較グライコーム研究会(6_05版).pdf

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発表場所：北海道 札幌市 札幌コンベンションセンター  

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発表場所：札幌コンベンションセンター、札幌市  

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その他リンク： http://www-user.yokohama-cu.ac.jp/~english/index.php/2020/06/11/a-ycu-alumnus-prof-dr-s-m-abe-kawsar-university-of-chittagong-bangladesh-published-two-academic-books-from-a-german-publisher/

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発表場所：インド農業科学大学 インド共和国、ベンガルール  

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発表場所：ベンガル―ル、インド共和国  

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発表場所：MDPI 出版 (スイス・バーゼル)  

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発表場所：横浜市立大 木原生物学研究所  

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発表場所：大さん橋ホール 横浜  

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発表場所：7日(木)、8日(金) 於お茶の水女子大  

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発表場所：秋葉原コンベンションホール５階カンファレンスフロア/５B会議室  

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発表場所：さくらWORKS〈関内〉イベントスペース  

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作品分類：教材  

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発表場所：横浜市立大学  

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作品分類：Web Service  

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発表場所：文部科学記者会・横浜市政記者クラブ  

2016年6月16日 日刊みなと新聞 掲載

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発表場所：横浜市立大学ウェッブサイト  

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発表場所：Croatia, Sprit  

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添付ファイル： 2014 - Ozeki - YCU, San Diego retreat program - SA_rev2.pdf

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発表場所：サンフォード・バーナム医学研究所、UCSD、スクリップス海洋研究所 他  

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発表場所：Orland, FL, USA  

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発表場所：横浜市立大学  

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発表場所：第22回国際複合糖質学会(Glyco22) 中国、大連  

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発表場所：海洋研究開発機構  

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発表場所：Protein Data Bank  

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作品分類：教材   発表場所：Yokohama City University  

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