INHIBITION OF CALPAIN IN INTACT PLATELETS BY THE THIOL PROTEASE INHIBITOR-E-64D
INHIBITION OF CALPAIN IN INTACT PLATELETS BY THE THIOL PROTEASE INHIBITOR-E-64D
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DOI：
10.1016/s0006-291x(89)80065-8
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发表时间：
1989-01-31
影响因子：
3.1
通讯作者：
DETWILER, TC
中科院分区：
文献类型：
--
作者：
MCGOWAN, EB;BECKER, E;DETWILER, TC
E-64d 是 E-64c 的膜渗透衍生物，E-64c 是一种硫醇蛋白酶抑制剂（Tamai 等人 (1986) J. Pharmacobio-Dyn. 9, 672-677），测试了抑制完整血小板中钙蛋白酶活性的能力。通过肌动蛋白结合蛋白和踝蛋白（钙蛋白酶的两种已知底物）的蛋白水解来测量钙蛋白酶活性。裂解前将血小板与 E-64c（不渗透）或 E-64d 一起孵育可防止裂解后的蛋白水解。当血小板与E-64c或E-64d一起孵育，然后在裂解前洗涤以除去药物时，只有E-64d抑制蛋白水解。当血小板与 E-64c 或 E-64d 一起孵育，然后用 A23187 加钙（一种激活血小板内钙蛋白酶的处理）激活时，只有 E-64d 抑制蛋白水解。这些结果表明E-64d可以进入完整细胞并抑制钙蛋白酶。
E-64d, a membrane permeant derivative of E-64c, a thiol protease inhibitor (Tamai et al. (1986) J. Pharmacobio-Dyn. 9, 672-677), was tested for ability to inhibit calpain activity in intact platelets. Calpain activity was measured by proteolysis of actin-binding protein and talin, two known substrates of calpain. Incubation of platelets with E-64c (not permeant) or E-64d before lysis prevented proteolysis after lysis. When the platelets were incubated with E-64c or E-64d and then washed to remove the drugs before lysis, only E-64d inhibited proteolysis. When platelets were incubated with E-64c or E-64d and then activated with A23187 plus calcium, a treatment tht activates intraplatelet calpain, only E-64d inhibited proteolysis. These results indicate that E-64d can enter the intact cell and inhibit calpain.