Carlson K E, Woolkalis M J, Newhouse M G, Manning D R
Mol Pharmacol. 1986 Nov;30(5):463-8.
Gs and Gi are guanine nucleotide-binding proteins that mediate the stimulation and inhibition, respectively, of adenylate cyclase. The extent to which the beta subunit common to these proteins may be associated with the cytoskeleton of S49 mouse lymphoma cells was assessed by procedures of differential detergent extraction and immunotransfer blotting. Treatment of cells with 1% Triton X-100 results in nearly quantitative extraction of cellular protein, phospholipid, tubulin, and the catalytic component of adenylate cyclase. Approximately 65% of the beta subunit is refractory to extraction. This population of the beta subunit, along with a population of actin presumed to originate from actin filaments, is subsequently solubilized with hypotonic medium containing 0.5% sodium deoxycholate and 1% Tween-40. The pattern of distribution of the beta subunit among sequential detergent extracts is corroborated by that of generated immunoreactive tryptic fragments. These results are consistent with the interaction of guanine nucleotide-binding proteins with the cytoskeleton.
Gs和Gi是鸟嘌呤核苷酸结合蛋白，分别介导对腺苷酸环化酶的刺激和抑制。通过差异去污剂提取和免疫印迹法评估了这些蛋白共有的β亚基与S49小鼠淋巴瘤细胞骨架相关的程度。用1% Triton X-100处理细胞会导致细胞蛋白、磷脂、微管蛋白和腺苷酸环化酶的催化成分几乎被定量提取。约65%的β亚基难以被提取。这部分β亚基以及推测源自肌动蛋白丝的一部分肌动蛋白随后用含有0.5%脱氧胆酸钠和1%吐温40的低渗介质溶解。β亚基在连续去污剂提取物中的分布模式与产生的免疫反应性胰蛋白酶片段的分布模式相符。这些结果与鸟嘌呤核苷酸结合蛋白与细胞骨架的相互作用一致。
