Effect of divalent cations on the limited proteolysis of prothrombin by thrombin.
Effect of divalent cations on the limited proteolysis of prothrombin by thrombin.
复制标题
二价阳离子对凝血酶对凝血酶原的有限蛋白水解的影响。
DOI：
10.1016/0003-9861(85)90067-0
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发表时间：
1985
影响因子：
3.9
通讯作者：
Tarvers,RC
中科院分区：
文献类型：
--
作者：
Church,FC;Lundblad,RL;Noyes,CM;Tarvers,RC
二价阳离子对牛α-凝血酶水解凝血酶原的抑制作用表现为Mn ~（2+）<$Ca ~（2+）<$Mg ~（2+）> Sr ~（2+）<$Ba ~（2+）。这种效果不是由于凝血酶的催化活性的抑制，如通过水解特定的合成底物，H-d-Phe-pipecolyl-Arg-p-nitroanilide（D-PhePipArgNA）所测量的。二价阳离子的存在不抑制γ-羧基谷氨酸（Gla）-结构域无凝血酶原的血栓蛋白水解。凝血酶原和Gladomainless凝血酶原被用作D-PhePipArgNA的血栓水解的竞争性抑制剂。计算的凝血酶原表观Ki值为18 μm。当Ca ~（2+）或Mn ~（2+）存在时，没有抑制作用。无Gla结构域凝血酶原的表观Ki值为28 μmin，无论是否存在Ca ~（2+）。添加二价阳离子的凝血酶原，但不Gladomainless凝血酶原，导致在改变蛋白质构象的高性能尺寸排阻色谱法和紫外差光谱测量。这些结果表明，二价阳离子与凝血酶原的含Gla结构域的相互作用的二级构象变化是需要阳离子依赖性抑制凝血酶水解。
The inhibitory influence of divalent cations on the ability of bovine α-thrombin to hydrolyze prothrombin showed the trend Mn2+⪢ Ca2+⩾ Mg2+> Sr2+⪢ Ba2+. This effect was not due to an inhibition of thrombin's catalytic activity as measured by hydrolysis of a specific synthetic substrate,H-d-Phe-pipecolyl-Arg-p-nitroanilide (D-PhePipArgNA). The presence of divalent cations did not inhibit thrombic proteolysis of γ-carboxyglutamic acid (Gla)-domainless prothrombin. Prothrombin and Gladomainless prothrombin were used as competitive inhibitors in the thrombic hydrolysis of D-PhePipArgNA. The apparentKivalue calculated for prothrombin was 18 μm. When either Ca2+or Mn2+were present, there was no inhibition. The apparentKivalue determined for Gla-domainless prothrombin was 28 μmin either the absence or presence of Ca2+. Addition of divalent cations to prothrombin, but not to Gladomainless prothrombin, resulted in an altered protein conformation as measured by high-performance size-exclusion chromatography and ultraviolet difference spectroscopy. These results suggest that a conformational change secondary to the interaction of divalent cations with the Gla-containing domain of prothrombin is required for cation-dependent inhibition of thrombin hydrolysis.