Yoshida K, Kimura H
Biochim Biophys Acta. 1986 Feb 19;880(2-3):197-203. doi: 10.1016/0304-4165(86)90080-2.
We have examined the effect of a mercurial sulfhydryl reagent, mersalyl, on the protein composition of cytoskeletons by SDS-polyacrylamide gel electrophoresis after treatment of human platelets with Triton X-100 (Triton) containing mersalyl and Ca2+, and have found that mersalyl alters the protein composition of cytoskeletons in a Ca2+-dependent manner. At 1 X 10(-7) M Ca2+, 0.2 mM mersalyl, which represents approximately the equivalent amount of sulfhydryl of platelet suspensions that we used, specifically made myosin insoluble. The amount of myosin in Triton-mersalyl residues was increased by increasing the Ca2+ concentration of Triton lysis buffer. Actin-binding protein, 235 kDa polypeptide and alpha-actinin-like protein were decreased in Triton residues by mersalyl at Ca2+ concentrations less than 1 X 10(-7) M, while these polypeptides in Triton residues were increased by mersalyl in the presence of more than 2 X 10(-7) M Ca2+. Electron microscopic study revealed the presence of thick filaments with an appearance similar to that of the thick filaments of platelet myosin. Thus, the modification with mersalyl of sulfhydryls of platelet polypeptides along with changes in Ca2+ concentrations within a physiological range leads to changes in solubility of, and filament formation of, myosin, actin and other cytoskeletal proteins.
我们在用含汞撒利和Ca2+的 Triton X-100（曲拉通）处理人血小板后，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳研究了汞巯基试剂汞撒利对细胞骨架蛋白质组成的影响，发现汞撒利以Ca2+依赖的方式改变细胞骨架的蛋白质组成。在1×10(-7) M Ca2+、0.2 mM汞撒利（约相当于我们所用血小板悬液中巯基的量）的条件下，汞撒利特异性地使肌球蛋白不溶。通过提高曲拉通裂解缓冲液中Ca2+的浓度，曲拉通-汞撒利残余物中肌球蛋白的量增加。在Ca2+浓度低于1×10(-7) M时，汞撒利使曲拉通残余物中的肌动蛋白结合蛋白、235 kDa多肽和α-辅肌动蛋白样蛋白减少，而在Ca2+浓度高于2×10(-7) M时，汞撒利使这些多肽增加。电子显微镜研究显示存在外观与血小板肌球蛋白粗丝相似的粗丝。因此，在生理范围内，随着Ca2+浓度的变化，汞撒利对血小板多肽巯基的修饰导致肌球蛋白、肌动蛋白和其他细胞骨架蛋白的溶解性和细丝形成发生变化。
