Weihing R R
J Cell Biol. 1976 Oct;71(1):303-7. doi: 10.1083/jcb.71.1.303.
When the 100,000 g supernatant fraction (extract) of HeLa cells lysed in a buffer containing sucrose, ATP, DTE, EGTA, imidazole, and Triton X-100 is incubated at 25 degrees C, it gels, and actin and a HMWP are progressively enriched in the extract and in gel isolated from extract. CB (greater than or equal to 0.25 muM) inhibits gelation and specifically lowers the concentrations of actin and the HMWP in the fraction which sediments at 100,000 g after incubation. These results indicate that actin and HMWP are partly disaggregated by cytochalasin treatment, and thus that their aggregation is related gelation. Inasmuch as previous results showed that actin is present and HMWP is enriched in the plasma membrane fraction of HeLa cells, the results also point to a possible relation between plasma membrane-associated gel and in vivo effects of CB.
当在含有蔗糖、ATP、二硫苏糖醇(DTE)、乙二醇双四乙酸(EGTA)、咪唑和曲拉通X - 100的缓冲液中裂解的HeLa细胞的100,000g上清液组分(提取物)在25℃下孵育时,它会凝胶化,并且肌动蛋白和一种高分子量蛋白(HMWP)在提取物以及从提取物中分离出的凝胶中逐渐富集。细胞松弛素B(CB,≥0.25μM)抑制凝胶化,并特异性降低孵育后在100,000g沉降的组分中肌动蛋白和HMWP的浓度。这些结果表明,细胞松弛素处理会使肌动蛋白和HMWP部分解聚,因此它们的聚集与凝胶化有关。鉴于先前的结果表明肌动蛋白存在且HMWP在HeLa细胞的质膜组分中富集,这些结果还表明质膜相关凝胶与CB的体内效应之间可能存在关联。