Hashimoto K, Tatsumi N
Biochem Int. 1986 May;12(5):653-60.
We investigated the effects of the N-ethylmaleimide (NEM), a sulfhydryl(SH) radical blocker, on platelet activation. Platelet aggregation and ATP release was suppressed by 0.2 mM NEM during ADP (20 microM) stimulation and by 0.5 mM NEM during A23187 (4 microM) stimulation. However the agent had no effect on actin polymerization in stimulated platelets. In the absence of a stimulant, NEM (over 1 mM) induced shape changes and slight (5%) actin polymerization, but not aggregation or ATP release. Although platelet aggregation and ATP release were suppressed by the addition of 1 mM NEM during the process of both reactions, the amount of polymerized actin was not influenced by the addition. The reconstructed system consisting of actin and partially purified regulatory proteins without myosin showed a dose-dependent increase in turbidity by the addition of NEM. From these findings, we concluded that NEM enhances actin polymerization, although actin molecules contain SH-radicals, and that actin polymerization has little affect on aggregation and release reaction.
我们研究了巯基(SH)自由基阻断剂N - 乙基马来酰亚胺(NEM)对血小板活化的影响。在ADP(20微摩尔)刺激期间,0.2毫摩尔的NEM可抑制血小板聚集和ATP释放;在A23187(4微摩尔)刺激期间,0.5毫摩尔的NEM可起到抑制作用。然而,该试剂对受刺激血小板中的肌动蛋白聚合没有影响。在没有刺激剂的情况下,NEM(超过1毫摩尔)可诱导形状改变和轻微(5%)的肌动蛋白聚合,但不会引起聚集或ATP释放。尽管在两个反应过程中添加1毫摩尔的NEM可抑制血小板聚集和ATP释放,但聚合肌动蛋白的量不受添加物的影响。由肌动蛋白和不含肌球蛋白的部分纯化调节蛋白组成的重建系统,通过添加NEM显示出剂量依赖性的浊度增加。从这些发现中,我们得出结论,尽管肌动蛋白分子含有SH自由基,但NEM可增强肌动蛋白聚合,并且肌动蛋白聚合对聚集和释放反应影响很小。