Development of cell surface linkage complexes in cultured fibroblasts.
Development of cell surface linkage complexes in cultured fibroblasts.
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培养的成纤维细胞中细胞表面连接复合物的发育。
DOI：
10.1083/jcb.100.4.1103
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发表时间：
1985-04
期刊：
影响因子：
--
通讯作者：
Yamada KM
中科院分区：
文献类型：
--
作者：
Chen WT;Hasegawa E;Hasegawa T;Weinstock C;Yamada KM
140 K膜相关蛋白复合物（140 K）在纤连蛋白-细胞骨架结合中的可能作用已被研究。用单克隆抗体JG 22 E鉴定140 K。单克隆抗体和多克隆抗体的140 K表现出相同的模式的固定和透化的鸡胚胎成纤维细胞的细胞膜的结合，本地化是弥漫性的，但在细胞与细胞外基质接触部位的显着浓度。干涉反射显微镜和双标记或三标记免疫荧光的相关定位显示，140 K与细胞外纤维连接蛋白原纤维和细胞内α-辅肌动蛋白共分布在细胞外基质接触部位的微丝束中，但不倾向于与原肌球蛋白共定位在远离粘附部位的部位的束中。此外，抗体与140 K、α-辅肌动蛋白和纤连蛋白的结合被排除在细胞外周的富含黏着斑蛋白的粘着斑位点之外。在早期扩散细胞中观察到细胞表面α-辅肌动蛋白-140K-纤连蛋白协会的逐步发展。抗140 K单克隆抗体JG 22 E抑制正常和劳斯肉瘤病毒转化的鸡胚成纤维细胞的纤维连接蛋白基质的附着和扩散。然而，抗140 K单克隆抗体成为一个积极的调解人的细胞附着和蔓延，如果它被吸附或交联的基质。我们的研究结果提供了第一个描述的膜相关蛋白复合物，共定位与纤连蛋白和微丝束，他们认为，140 K复合物可能是一个细胞表面的纤连蛋白和细胞骨架之间的联系。
The possible role of a 140K membrane-associated protein complex (140K) in fibronectin-cytoskeleton associations has been examined. The 140K was identified by the monoclonal antibody JG22E. Monoclonal and polyclonal antibodies to the 140K showed identical patterns of binding to the cell membranes of fixed and permeabilized chicken embryonic fibroblasts; localization was diffuse, but with marked concentration in cell-to-extracellular matrix contact sites. Correlative localization with interference reflection microscopy and double-label or triple- label immunofluorescence showed that 140K co-distributed with extracellular fibronectin fibrils and intracellular alpha-actinin in microfilament bundles at extracellular matrix contact sites but tended not to co-localize with tropomyosin present in bundles at sites farther from adhesion sites. In addition, binding of antibodies to 140K, alpha- actinin, and fibronectin was excluded from vinculin-rich focal adhesion sites at the cellular periphery. A progressive development of cell surface alpha-actinin-140K-fibronectin associations was observed in early spreading cells. The anti-140K monoclonal antibody JG22E inhibited the attachment and spreading of both normal and Rous sarcoma virus-transformed chicken embryonic fibroblasts to a fibronectin substratum. However, the anti-140K monoclonal antibody became a positive mediator of cell attachment and spreading if it was adsorbed or cross-linked to the substratum. Our results provide the first description of a membrane-associated protein complex that co-localizes with fibronectin and microfilament bundles, and they suggest that the 140K complex may be part of a cell surface linkage between fibronectin and the cytoskeleton.