Abstract：
Thrombosis is characterized by the formation of clots in the blood vessels. Antithrombin-III deficiency in the blood causes thrombus formation. Supplementing antithrombin-III may serve as anticoagulant therapy. In the present studies, an antithrombin like Protein from Punica granatum has been identified and characterized using in silico approach. Based on sequence homology, an ALPP was selected depending upon its highest binding affinity of -41.28 kcal/mol with thrombin. Thrombin structure complexed with ALPP was docked with TAME using AutoDock Vina. No binding was observed for TAME at Ser195 of thrombin. MD simulation (50 ns) was performed to evaluate the flexibility and stability of docked complexes. In vitro assays with crude protein showed 78% thrombin inhibition at 5 µg and calculated IC50 value was 0.188 µg. The presence of thrombin inhibitors in crude protein was also confirmed by reverse zymography. Thus, it is very likely that the protein identified from P. granatum may act as thrombin inhibitor.
摘要：
血栓形成的特征在于在血管中形成凝块。血液中的抗凝血酶III缺乏导致血栓形成。补充抗凝血酶-III可以作为抗凝治疗。在目前的研究中,已使用计算机模拟方法鉴定和表征了来自石榴石的抗凝血酶样蛋白。基于序列同源性,根据其与凝血酶的最高结合亲和力-41.28kcal/mol来选择ALPP。使用AutoDockVina将与ALPP复合的凝血酶结构与TAME对接。在凝血酶的Ser195处没有观察到TAME的结合。进行MD模拟（50ns）以评估对接复合物的柔韧性和稳定性。粗蛋白的体外测定显示，在5μg时，凝血酶抑制为78％，计算的IC50值为0.188μg。粗蛋白中凝血酶抑制剂的存在也通过反向酶谱证实。因此，很可能从石榴中鉴定出的蛋白质可作为凝血酶抑制剂。
