| Project/Area Number |
04671378
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Biological pharmacy
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| Research Institution | Kyoto Sangyo University |
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Principal Investigator |
NAKADA Hiroshi Department of Biotechnology, Kyoto Sangyo University, Associate Professor, 工学部・生物工学科, 助教授 (90113141)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1993: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1992: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Keywords | Tn antigen / Glycophorin A / Leulosialin / Sialyl-Le^a / T抗原 / ムチン |
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| Research Abstract |
Expression and strucutre of Tn antigen was investigated by using human glycophorin A and leulosialin. Glycophorin A was treated with glycoprotease and the digest was fractionated by various volumn chromatographies. Several glycopeptides tith a cluster and /or single structure on an O-glycan were obtained, which were certified by chemical and sequence analyzes. Tn antigenecty of these glycopeptoeds were examined by solid phasa radioimmunoassay after treatment with sialidase and-galactosidase. Essential structure of Tn antigenecity was found to be three consecutive glycosylated Ser/Thr residues. A large amount of leukosialin was also prepared form human T cell lymphoma, Jurkat cells and the digests with trypsin was applied to a immunoaffinity column of MLS128 (anti Tn antibody). Immunoreactive glycopeptides thus obtained were purified by reverse phase chromatography. Sequence analyzes revealed that all the glycopeptieds obtained contained three consequtive residues ofM-acetylgalactosamine-Ser/Thr. Novel gangliosides containing sialyl-Le^a were also found in a juman rectal adenocarcinoma. Oligosaccharides were released from the gangliosides of a human rectal adenocarcinoma with endoglycoceramidase and then purifies by affinity chromatography ona column of an immobilized monoclonal antibody, MSW113. Structural studies, involving 600-MHz-^1HNMR spectrometry indicated that umique lkigosaccharide chains containing the sialyl-Le^a-X antigen in linear and brached structures were found, which might be new potential tumar markers and/or ligands for selectin families.
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