INVESTIGATION OF THE ROLE OF CALPAIN AS A STIMULUS-RESPONSE MEDIATOR IN HUMAN PLATELETS USING NEW SYNTHETIC INHIBITORS
INVESTIGATION OF THE ROLE OF CALPAIN AS A STIMULUS-RESPONSE MEDIATOR IN HUMAN PLATELETS USING NEW SYNTHETIC INHIBITORS
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DOI：
10.1042/bj2740497
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发表时间：
1991-03-01
影响因子：
4.1
通讯作者：
SHAW, E
中科院分区：
文献类型：
--
作者：
ANAGLI, J;HAGMANN, J;SHAW, E
合成了一系列具有钙蛋白酶特异性结构的肽基重氮甲烷和一氟甲烷，并以人血小板为模型系统，在体内测试了它们抑制钙蛋白酶活性的能力。通过观察肌动蛋白结合蛋白和Talin这两种已知的钙蛋白酶底物的蛋白分解来确定体内钙蛋白酶的活性。从这项研究中出现的非常有效的抑制剂被用来研究钙蛋白酶在一些血小板反应过程中的作用。我们的结果表明，钙蛋白酶介导的血小板蛋白分解不是导致细胞形状改变、与玻璃黏附和铺展、聚集和5-羟色胺释放的必备事件。其中两种抑制剂被I-125碘标记，并在体内用于放射性标记酶。据我们所知，这项工作也是第一份报告，描述了使用不可逆转的放射性抑制剂亲和标记人血小板中的钙蛋白酶。
A series of peptidyl diazomethanes and monofluoromethane with structures specific for calpain have been synthesized and tested for their ability to inhibit calpain activity in vivo, using human platelets as a model system. Calpain activity in vivo was determined by observing proteolysis of actin-binding protein and talin, two known substrates of calpain. Very potent inhibitors, which emerged from this study, were used to investigate the role of calpain in some platelet response processes. Our results show that calpain-mediated proteolysis in platelets is not an obligatory event leading to change of cell shape, adhesion to glass and spreading, aggregation and 5-hydroxytryptamine release. Two of the inhibitors were iodinated with I-125 and used to radiolabel the enzyme in vivo. To our knowledge, this work also represents the first report describing the affinity labelling of calpain in human platelets using irreversible radioactive inhibitors.