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Function and expression mechanism of cancer associated carbohydrate antigens

癌症相关糖抗原的功能和表达机制

基本信息

批准号：
04671378
负责人：
NAKADA Hiroshi
金额：
$ 1.34万
依托单位：
Kyoto Sangyo University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1992
资助国家：
日本
起止时间：
1992 至 1993
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04671378/
关键词：
Tn antigen Glycophorin A Leulosialin Sialyl-Le^a T抗原ムチン

项目摘要

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.

用人血型糖蛋白A和leulosialin研究Tn抗原的表达和结构。用糖蛋白酶处理血型糖蛋白A，并通过各种体积色谱法对消化物进行分级。通过化学分析和序列分析，得到了几个在O-聚糖上具有簇和/或单一结构的糖肽。用唾液酸酶和β-半乳糖苷酶处理后，用固相放射免疫分析法检测这些糖肽的抗原性。发现Tn抗原性的基本结构是三个连续的糖基化Ser/Thr残基。从人T细胞淋巴瘤、Jurkat细胞中提取大量唾液酸白蛋白，用胰蛋白酶消化，用MLS 128（抗Tn抗体）免疫亲和柱进行亲和层析。通过反相色谱法纯化由此获得的免疫反应性糖肽。序列分析表明，所获得的糖肽均含有M-乙酰半乳糖胺-Ser/Thr的3个连续残基。在juman直肠腺癌中也发现了含有唾液酸-Le ^a的新型神经节苷脂。用神经酰胺内切酶从人直肠腺癌神经节苷脂中释放寡聚腺苷酸，然后用固定化单克隆抗体MSW 113柱进行亲和层析纯化。利用600-MHz-^1HNMR谱进行的结构研究表明，发现了含有唾液酸-Le ^a-X抗原的线性和分支结构的独特寡糖链，它们可能是选择素家族的新的潜在肿瘤标记物和/或配体。