The hyaluronate receptor is associated with actin filaments.
The hyaluronate receptor is associated with actin filaments.
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透明质酸盐受体与肌动蛋白丝有关。
DOI：
10.1083/jcb.105.3.1395
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发表时间：
1987-09
影响因子：
7.8
通讯作者：
Underhill, C B
中科院分区：
文献类型：
--
作者：
Lacy, B E;Underhill, C B
透明质酸的细胞表面受体是一种完整的膜糖蛋白，分子量为85,000(UnderHill，C.B.，A.L.瑟恩和B.E.Lacy，1985，J.Biol.化学，260：8128-8133)，它被认为介导了许多透明质酸对细胞行为的影响，如迁移、血管生成和吞噬作用。为了确定受体是否与潜在的细胞骨架相关，用Triton X-100溶液提取瑞士3T3细胞，Triton X-100溶液溶解了大部分细胞成分，但留下了含有细胞骨架的不溶残留物。这种不溶于洗涤剂的残基被发现含有大量的透明质酸结合活性，这表明该受体可能确实与细胞骨架有关。为了进一步确定与受体相互作用的细胞骨架元件，用Triton X-100在各种不同的离子条件下提取了3T3细胞。在每种情况下，不溶于洗涤剂的残留物中的透明质酸结合活性与肌动蛋白的量有关，但与微管蛋白或波形蛋白无关。此外，如果细胞在鬼臼毒素(一种稳定肌动蛋白细丝的药物)存在的情况下提取，透明质酸结合活性的恢复显著增强(在大多数情况下达到100%)。然而，当整个细胞在提取前用细胞松弛素B处理时，结合活性的恢复显著降低，当提取的细胞用促进肌动蛋白细丝解聚的DNase I处理时，结合活性的恢复显著降低。此外，将SV-40转化的瑞士3T3细胞膜的提取物与DNase I预先孵育后，分子筛层析判断，受体的洗脱曲线发生了变化。据推测，受体大小的减少是由于相关肌动蛋白细丝的丢失。这些实验的结果强烈地表明，透明质酸受体与胞浆肌动蛋白细丝直接或间接相关。
The cell-surface receptor for hyaluronate is an integral membrane glycoprotein of Mr 85,000 (Underhill, C. B., A. L. Thurn, and B. E. Lacy, 1985, J. Biol. Chem., 260:8128-8133) that is thought to mediate many of the effects that hyaluronate has on cell behavior, such as migration, angiogenesis, and phagocytosis. To determine if the receptor is associated with the underlying cytoskeleton, Swiss 3T3 cells were extracted with a solution of Triton X-100, which solubilized most of the cellular components, but which left behind an insoluble residue containing the cytoskeleton. This detergent-insoluble residue was found to contain the bulk of the hyaluronate-binding activity, suggesting that the receptor might indeed be associated with the cytoskeleton. To further define the cytoskeletal element with which the receptor interacts, 3T3 cells were extracted with Triton X-100 under a variety of different ionic conditions. In each case, the amount of hyaluronate- binding activity in the detergent-insoluble residue was related to the amount of actin present, but not to either tubulin or vimentin. In addition, the recovery of hyaluronate-binding activity was dramatically enhanced (to 100% in most cases) if the cells were extracted in the presence of phalloidin, a drug that stabilizes actin filaments. However, the recovery of binding activity was dramatically decreased when whole cells were treated with cytochalasin B before extraction, and when extracted cells were treated with DNase I, which promotes the depolymerization of actin filaments. In addition, preincubating an extract of SV-40-transformed Swiss 3T3 cell membranes with DNase I caused a change in the elution profile of the receptor as judged by molecular-sieve chromatography. Presumably this decrease in the size of the receptor is due to the loss of associated actin filaments. The results of these experiments strongly suggest that the receptor for hyaluronate is associated either directly or indirectly with cytosolic actin filaments.