肝脏中微管相关蛋白的研究——从兔肝脏中分离出一种210 kDa的微管相关蛋白

[Studies of microtubule associated proteins in the liver--isolation of a 210 kDa microtubule associated protein from rabbit liver].

作者信息

Komatsu M, Yamamoto A, Goto M, Tobori F, Yagisawa H, Toyoshima I, Masamune O

出版信息

Nihon Shokakibyo Gakkai Zasshi. 1989 Dec;86(12):2773-8.

Abstract

The biochemical characteristics and cellular localization of microtubule associated proteins (MAPs) were studied using rabbit liver. The MAPs fraction, isolated from rabbit liver crude extract by a modification of Vallee's method, consisted of several proteins, as identified by SDS-polyacrylamide gel electrophoresis. The most prominent of these was a 210 kDa protein (rabbit 210 kDa MAP). The molecular weight of rabbit 210 kDa MAP was distinct from that of the major brain MAPs, MAP1, MAP2 and Tau, isolated from rabbit brain crude extract by same method. Monoclonal antibodies (mAb) to this rabbit 210 kDa MAP were raised. Using immunoblotting method, we confirmed that anti-rabbit 210 kDa MAP mAb reacted to 210 kDa protein from rabbit liver excrude, while this mAb also reacted to 210 kDa protein from rabbit brain excrude. Immunofluorescent staining using fixed rabbit liver was carried out using these mAb. In frozen sections of the rabbit liver, the antigen related to mAb 7B10 was localised in the hepatic cells and epithelial cells of the bile duct but not in the smooth muscle cells and endothelial cells.

摘要

利用兔肝研究了微管相关蛋白（MAPs）的生化特性和细胞定位。通过对瓦利方法的改进，从兔肝粗提物中分离得到的MAPs组分由几种蛋白质组成，这是通过SDS-聚丙烯酰胺凝胶电泳鉴定的。其中最突出的是一种210 kDa的蛋白质（兔210 kDa MAP）。兔210 kDa MAP的分子量与通过相同方法从兔脑粗提物中分离得到的主要脑MAPs，即MAP1、MAP2和Tau不同。制备了针对这种兔210 kDa MAP的单克隆抗体（mAb）。使用免疫印迹法，我们证实抗兔210 kDa MAP mAb与兔肝粗提物中的210 kDa蛋白质反应，而该mAb也与兔脑粗提物中的210 kDa蛋白质反应。使用这些mAb对固定的兔肝进行免疫荧光染色。在兔肝的冰冻切片中，与mAb 7B10相关的抗原定位于肝细胞和胆管上皮细胞中，而不在平滑肌细胞和内皮细胞中。