Volk T, Geiger B
EMBO J. 1984 Oct;3(10):2249-60. doi: 10.1002/j.1460-2075.1984.tb02123.x.
We report here on a new 135-kd membrane protein which is specifically associated with intercellular adherens-type junctions. This surface component was identified by a monoclonal antibody, ID-7.2.3, raised against detergent-extracted components of membranes of chicken cardiac muscle rich in intercalated discs. The antibodies stain extensively adherens junctions in intact cardiac muscle and in lens, as well as in cultured cells derived from these tissues. In living cultured cells only very little immunolabelling was obtained with ID-7.2.3 antibodies, probably due to the limited accessibility of the antibodies to the intercellular gap. However, upon the removal of extracellular Ca2+ ions a dissociation of the junction occurred, leading to the rapid exposure of the 135-kd protein. Immunoelectron microscopic labelling of EGTA-treated, or detergent-permeabilized cells indicated that the antigen is found along the plasma membrane and highly enriched in contact areas. Double immunolabelling for both the 135-kd protein and vinculin pointed to the close association of the two in intercellular junctions and to the apparent absence of the former protein from the vinculin-rich focal contacts of cultured cells and from dense plaque of smooth muscle. Immunoblotting indicated that the 135-kd protein is present in many tissues but is particularly enriched in heart, lens and brain.
我们在此报告一种新的135千道尔顿膜蛋白,它与细胞间黏附型连接特别相关。这种表面成分是由一种单克隆抗体ID-7.2.3鉴定出来的,该抗体是针对富含闰盘的鸡心肌膜的去污剂提取物产生的。这些抗体能广泛标记完整心肌、晶状体以及源自这些组织的培养细胞中的黏附连接。在活的培养细胞中,用ID-7.2.3抗体仅获得很少的免疫标记,这可能是由于抗体进入细胞间隙的能力有限。然而,去除细胞外钙离子后,连接会发生解离,导致135千道尔顿蛋白迅速暴露。对经乙二醇双乙酸盐(EGTA)处理或去污剂通透处理的细胞进行免疫电子显微镜标记表明,该抗原沿质膜分布,且在接触区域高度富集。对135千道尔顿蛋白和纽蛋白进行双重免疫标记表明,二者在细胞间连接中紧密相关,且在培养细胞富含纽蛋白的粘着斑和平滑肌致密斑中明显不存在前一种蛋白。免疫印迹表明,135千道尔顿蛋白存在于许多组织中,但在心脏、晶状体和大脑中尤其丰富。