Expression of catabolic pathway of asparagine-linked oligosaccharides on plasma membrane and apoptosis

天冬酰胺连接寡糖分解代谢途径在质膜上的表达与细胞凋亡

基本信息

批准号：
14580628
负责人：
ITO Kazuo
金额：
$ 2.24万
依托单位：
Osaka City University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

项目摘要

1.Verification and specification of target glycoproteins deglycosylated by Endo HSTarget glycoprotein(TGP) deglycosylated by Endo HS was detected from attached epithelial cell in the human oral cavity more than detached cell, but not in leukocyte. The protein of molecular weight(MW) of 55K from the attached epithelial cell was specified as TGP55 by in-gel deglycosylation method. The proteins of MW of 20K, 50K and 105〜110K were also specified as TGP20, TGP50 and TGP105〜110.2.Identification of TGP55 and TGP50TGP55 and TGP50 were purified. The N-terminal amino acid was thought to be acetylated. The N-terminal amino acid sequence of deacetylated TGP55 showed high homology with the inner amino acid sequence of human cytokeratin 13, but no proteins having the same N-terminal amino acid sequence was not observed by homology search. The N-terminal amino acid of TGP50 was also blocked. The N-terminal and inner amino acid sequence of TGP50 showed high homology with those of human type I cytokerain 13, indicating that TGP50 is type I cytokeratin 13 or a similar protein to it. Human type I cytokeratin 13 has an asparagine of potential glycosylation site. Endo HS was thought to removing the asparagine-linked oligosaccharide at the site and taking part in the differentiation of the epithelial cell.3.Immunochemical properties and cellular localization of TGP55 analyzed using monoclonal antibodies against TGP55Three monoclonal antibodies against TGP55 were prepared. Each antibody reacted with TGP55 and other proteins of attached and detached epithelial cell, the kinds of proteins were different between attached and detached epithelial cell. The outskirts part of the epithelial cell was stained by the antibodies, indicating that TGP55 localizes around the outskirt part of the epithelial cell and the localization changes after peeling of the epithelial cell.

1.验证和规范通过内核糖基化糖基化糖基化的糖基因糖基糖基糖基化的糖基因糖基糖基化（TGP）通过内部型HS糖基化的糖基化糖基于人类口腔中的上皮细胞被检测到比分离的细胞中的上皮细胞，但在白细胞中没有检测到。通过凝胶脱胶糖基化方法将来自附着的上皮细胞的55K分子量（MW）的蛋白质（MW）指定为TGP55。还指定为20K，50K和105-110K的MW的蛋白质为TGP20，TGP50和TGP105-110.2。净化TGP55和TGP50TGP55和TGP50的识别。脱乙酰化的N末端氨基酸序列。脱乙酰化TGP55的N末端氨基酸序列与人细胞角蛋白13的内部氨基酸序列具有很高的同源性，但没有通过同源性搜索观察到具有相同N末端氨基酸序列的蛋白质。 TGP50的N末端氨基酸也被阻塞。 TGP50的N末端和内部氨基酸序列与人类I型胞质量13的同源性很高，表明TGP50是I型细胞角因13或类似蛋白的同源性。人类I型胞质因子13具有潜在糖基化位点的天冬酰胺。人们认为Endo HS可以去除现场的天冬酰胺连接的寡糖，并参与上皮细胞的分化。3。使用单克隆抗体对TGP55针对TGP55的TGP55分析的TGP55的免疫化学特性和细胞定位，对TGP55进行了针对TGP55的TGP55的TGP55。每种抗体与TGP55和附着和分离的上皮细胞的其他蛋白质反应，蛋白质的种类在附着和分离的上皮细胞之间不同。上皮细胞的郊区部分被抗体染色，表明TGP55位于上皮细胞郊区的部分周围，并且上皮细胞剥离后的定位变化。