Jander R, Troyer D, Rauterberg J
Biochemistry. 1984 Jul 31;23(16):3675-81. doi: 10.1021/bi00311a016.
The 140 000-dalton collagenous glycoprotein (CGP) from calf aorta and ligament characterized by Gibson & Cleary (1982) [Gibson, M.A., & Cleary, E.G. (1982) Biochem. Biophys. Res. Commun. 105, 1288-1295] has been studied. In the electron microscope, rotary-shadowed CGP molecules appear similar to the dimers of type VI collagen (short-chain collagen, intima collagen) described by other authors [Furthmayr, H., Wiedemann, H., Timpl, R., Odermatt, E., & Engel, J. (1983) Biochem. J. 211, 303-311] except that they have larger globular domains. As shown by gel electrophoresis, pepsin treatment of CGP at 4 degrees C either before or after reduction releases polypeptide chains corresponding in size to those of type VI collagen. Electron microscopic examination shows that pepsin digestion of nonreduced CGP removes the outer globular domains, reduces the size of the inner ones, and separates the paired central strands. The residual structures look like type VI collagen dimers. When intact CGP is reduced, monomers with two large globular ends are obtained. Pepsin digestion of monomers removes most or all of both globular domains. In immunoblots, CGP and its pepsin-derived fragments react with antibodies directed against type VI collagen. The results indicate that type VI collagen is an integral component of CGP.
对由吉布森和克利里(1982年)[吉布森,M.A.,& 克利里,E.G.(1982年)《生物化学与生物物理研究通讯》105,1288 - 1295]所描述的来自小牛主动脉和韧带的140000道尔顿胶原糖蛋白(CGP)进行了研究。在电子显微镜下,旋转阴影处理后的CGP分子看起来与其他作者所描述的VI型胶原(短链胶原、内膜胶原)二聚体相似[弗特迈尔,H.,维德曼,H.,蒂姆普尔,R.,奥德马特,E.,& 恩格尔,J.(1983年)《生物化学杂志》211,303 - 311],只是它们具有更大的球状结构域。如凝胶电泳所示,在4℃下对CGP进行还原前后用胃蛋白酶处理,会释放出大小与VI型胶原多肽链相对应的多肽链。电子显微镜检查表明,对未还原的CGP进行胃蛋白酶消化会去除外部球状结构域,减小内部球状结构域的大小,并分离出成对的中央链。剩余结构看起来像VI型胶原二聚体。当完整的CGP被还原时,会得到带有两个大球状末端的单体。对单体进行胃蛋白酶消化会去除大部分或所有球状结构域。在免疫印迹中,CGP及其胃蛋白酶衍生片段与针对VI型胶原的抗体发生反应。结果表明,VI型胶原是CGP的一个组成部分。