| Project/Area Number |
14572054
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biological pharmacy
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
KAWASAKI Nobuko Kyoto University, Faculty of Medicine, Professor, 医学部, 教授 (70077676)
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| Co-Investigator(Kenkyū-buntansha) |
KAWASAKI Toshisuke Kyoto University, Graduate School of Pharmaceutical Sciences, Professor, 薬学研究科, 教授 (50025706)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2003: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2002: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | serum lectin / mannan-binding protein / mannose-binding protein / human colon cancer cell / anti-tumor activity / host defence factor / oligosaccharide ligand / polylactosamine structure / ヒト結腸ガン細胞 / 赤血球Band3糖タンパク質 |
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| Research Abstract |
Serum Mannan-binding protein (MBP), a C-type mammalian lectin specific for mannose, N-acetylglucosamine and fucose, has a potent growth inhibitory activity to a human colorectal carcinoma cell line in vivo. In this study, we have characterized the MBP-ligand oligosaccharides expressed on the human colorectal carcinoma cell line, SW 1116, which are suggested to trigger anti-tumor activity of MBP.
1.FITC-labeled MBP binding to SW1116 cells was inhibited specifically by the ligand sugars, a fucose-specific lectin, and anti-Lewis A and anti-Lewis B mAbs, suggesting that MBP binds to type-I carbohydrates.
2.MBP-ligand glycopeptides were isolated by a MBP affinity column from the high molecular weight glycopeptides fraction of the pronase-digest of SW 1116 cell lysates. After hydrazinolysis followed by pyridylamination of MBP-ligand glycopeptides, PA-derivatized MBP-ligand oligosaccharides were isolated again by the MBP affinity column.
(1)Carbohydrate analysis indicated that MBP-ligand oligosaccharides contained high molecular size N-glycans with high galactose, N-acetylglucosamine and fucose contents.
(2)Endo-β-galactosidase digestion of the MBP-ligand oligosaccharides resulted in a marked reduction of the binding activity to the MBP column together with the decrease of their molecular sizes, indicating the presence of poly N-acetyllactosamine structures. The selective removal of fucose residues from MBP-ligand oligosaccharides resulted in almost complete loss of the binding activity to the MBP and the AAL affinity columns.
(3)MALDI-MSIMS and nano ESI-MS/MS analyses of the MBP-ligand oligosaccharides indicated the presence of highly fucosylated structure.
MBP-ligand oligosaccharides appear to be high molecular weight poly N-acetyllactosamine-type with high fucose content.
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