STRUCTURE AND FUNCTION OF A NOVEL BASEMENT MEMBRANE-ASSOCIATED COLLAGEN

新型基底膜相关胶原蛋白的结构和功能

基本信息

批准号：
03454538
负责人：
HAYASHI Toshihiko
金额：
$ 0.64万
依托单位：
THE UNIVERSITY OF TOKYO
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1991
资助国家：
日本
起止时间：
1991 至 1992
项目状态：
已结题

项目摘要

The novel collagen in the present project was defined as a protein specifically recognized by a monoclonal antibody originally developed for type IV collagen fraction from human placenta by pepsinization. The epitope appeared to reside on a polypeptide which contained an unidetified amino acid sequence. The collagen was found to be associated with a restricted basement membrane in the immunohistochemical examination on a vascular basement membrane and at least a portion of the polypeptide was associated with alpha 1(IV) chain, since a sandwith ELISA system using the other monoclonal antibody which recognizes alpha 1(IV) showed a positive reaction to the type IV collagen fraction, while the reaction became negative by treatment with heat or/and SH reagent. Molecular structure and function of the novel collagen was characterized by comparison with other extracellular components in solubility, intact size of the polypeptide, susceptibility to proteases. The study indicated similarity to an alpha chain of type IV collagen; that is 180kD chain was recovered only after treatment with SH reagent and denaturants from human placenta. The monoclonal antibody crossed with a particular sequence of alpha 1(IV) chain weakly. An important observation was that human fetal lung fibroblasts produced and deposited the novel collagen as well as alpha 1(IV) on the cell layer by culture, although human dermal fibroblasts deposited the collagen to a much less quantity from the study using cell-layer ELISA. Immunohistochemical localization indicated that deposition of novel collagen appeared to increase by fibroses of liver or kidney. Serum levels of the complex of the novel collagen as determined by ELISA were increased in patients with liver cirrhosis.

本项目中的新型胶原蛋白被定义为一种蛋白质，该蛋白质是由最初通过胃启动从人体胎盘中开发的单克隆抗体特异性识别的。表位似乎驻留在包含未经检测的氨基酸序列的多肽上。发现该胶原蛋白与在血管基底膜上的免疫组织化学检查中与受限的地下膜有关，至少部分多肽与α1（iv）链有关，因为使用夹层ELISA系统使用其他单克隆抗体进行了反应，而鉴定了Alpha 1（IV），这是鉴定的，IV（IV）conter conter conter conter conter conter conter conter conter anpha collpha 1（IV）。或/和SH试剂。新型胶原蛋白的分子结构和功能的特征是与其他细胞外成分相比，在溶解度，多肽的完整大小，对蛋白酶的敏感性中。该研究表明与IV型胶原蛋白的α链相似。那是只有在用SH试剂和人胎盘变性剂处理后才能回收180kD链。单克隆抗体与特定的α1（iv）链的特定序列较弱。一个重要的观察结果是，人类胎儿肺成纤维细胞通过培养产生并沉积了新颖的胶原蛋白以及α1（iv），尽管人类真皮成纤维细胞使用细胞层ELISA沉积了胶原蛋白的人类真皮成纤维细胞的数量。免疫组织化学定位表明，新型胶原蛋白的沉积似乎通过肝脏或肾脏的纤维蛋白增加。在肝硬化患者中，由ELISA确定的新型胶原蛋白复合物的血清水平升高。