Bourguignon L Y, Walker G, Suchard S J, Balazovich K
J Cell Biol. 1986 Jun;102(6):2115-24. doi: 10.1083/jcb.102.6.2115.
In this study we have used several complementary techniques to isolate and characterize a 72-kD polypeptide that is tightly associated with a major mouse T-lymphoma membrane glycoprotein, gp 85 (a wheat germ agglutinin-binding protein), in a 16 S complex. These two proteins do not separate in the presence of high salt but can be dissociated by treatment with 2 M urea. Further analysis indicates that the 72-kD protein has ankyrin-like properties based on the following criteria: (a) it cross-reacts with specific antibodies raised against erythrocyte and brain ankyrin; (b) it displays a peptide mapping pattern and a pI (between 6.5 and 6.8) similar to that of the 72-kD proteolytic fragment of erythrocyte ankyrin; (c) it competes with erythrocyte ghost membranes (spectrin-depleted preparations) for spectrin binding; and (d) it binds to purified spectrin and fodrin molecules. Most importantly, in intact lymphoma cells this ankyrin-like protein is localized directly underneath the plasma membrane and is found to be preferentially accumulated beneath receptor cap structures as well as associated with a membrane-cytoskeleton complex preparation. It is proposed that the ankyrin-like 72-kD protein may play an important role in linking certain surface glycoprotein(s) to fodrin which, in turn, binds to actin filaments required for lymphocyte cap formation.
在本研究中,我们运用了多种互补技术来分离和鉴定一种72-kD多肽,该多肽在一个16S复合物中与小鼠主要的T淋巴瘤膜糖蛋白gp 85(一种小麦胚凝集素结合蛋白)紧密相连。在高盐存在的情况下,这两种蛋白不会分离,但用2M尿素处理后可使其解离。进一步分析表明,基于以下标准,该72-kD蛋白具有锚蛋白样特性:(a)它能与针对红细胞和脑锚蛋白产生的特异性抗体发生交叉反应;(b)它呈现出与红细胞锚蛋白的72-kD蛋白水解片段相似的肽图谱和pI(在6.5至6.8之间);(c)它与红细胞血影膜(血影蛋白缺失制剂)竞争血影蛋白结合;(d)它能与纯化的血影蛋白和细丝蛋白分子结合。最重要的是,在完整的淋巴瘤细胞中,这种锚蛋白样蛋白直接定位在质膜下方,并且发现它优先聚集在受体帽结构下方,同时与一种膜-细胞骨架复合物制剂相关联。有人提出,这种72-kD的锚蛋白样蛋白可能在将某些表面糖蛋白与细丝蛋白连接方面发挥重要作用,而细丝蛋白又与淋巴细胞帽形成所需的肌动蛋白丝结合。