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本实验研究蛇毒蛋白C激活组份F6.3.2对蛋白C的激活作用。以BaCl2吸附血浆和纯化蛋白C为作用底物,采用发色底物法测定蛇毒组份F6.3.2对蛋白C的特异性激活作用,同时采用SDS-PAGE.测定该组份对蛋白C的激活机制。结果发现F6.3.2没有直接作用发色底物使之生色的能力。它对BaCl2吸附血浆不表现生色反应能力(与对照组相比P>0.05),而对纯化蛋白C则表现出很强的生色反应能力(与对照组Ⅰ、对照组Ⅱ相比P<0.01).SDS-PAGE结果表明,F6.3.2与纯化蛋白C作用后的混合物,由两条链组成,一条链分子量为19KDa,与纯化蛋白C的轻链相同,另一条链分子量为59.5 KDa。为纯化蛋白C重链与F6.3.2的分子量之和,表明F6.3.2已结合到纯化蛋白C的重链上。因此.我们认为蛇毒蛋白C激活组份F6.3.2对蛋白C有特异性激活作用,它激活蛋白C的可能机制是通过结合于蛋白C的重链,形成F6.3.2-蛋白C复合物,引起蛋白C变构,从而将蛋白C激活为活化蛋白C。
This experiment investigated the activation of protein C by the venom protein C activating fraction F6.3.2. Using BaCl2 as adsorbent plasma and purified protein C as substrates, a chromogenic substrate method was used to determine the specific activation of protein C by the venom component F6.3.2, and SDS-PAGE was used to determine the activation of protein C by this component. mechanism. As a result, it was found that F6.3.2 had no direct effect on the ability of the chromogenic substrate to color. It showed no chromogenic response to BaCl2 adsorbed plasma (P>0.05 compared with control group), but showed strong chromogenic response to purified protein C (Compared to control group I and control group II, P< 0.01). SDS-PAGE results showed that the mixture of F6.3.2 and purified protein C consisted of two chains, one with a molecular weight of 19 KDa, the same as the light chain of purified protein C, and the other with a molecular weight of 59.5 KDa. In order to purify the sum of the molecular weights of protein C heavy chain and F6.3.2, it was shown that F6.3.2 has bound to the heavy chain of purified protein C. Therefore, we believe that the activated component of venom protein C, F6.3.2, specifically activates protein C. Its possible mechanism for activating protein C is to bind to the heavy chain of protein C and form a F6.3.2-protein C complex. Protein C is allosterically, so that protein C is activated as activating protein C.