4-甲酰苯基硼酸对蛋白酶MP的抑制作用及其抑制机制的分子动力学模拟  

Molecular Dynamical Simulation of Inhibitory Effects and Mechanism of 4-Formyl-Phenyl-Boronic Acid on Marine Protease

作　　者：纪晓峰[1] 郑媛[1] 王致鹏[1] 盛军[1] 王海英[1] JI Xiao-Feng;ZHENG Yuan;WANG Zhi-Peng;SHENG Jun;WANG Hai-Ying(Laboratory of Marine Enzyme and Engineering, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, Shandong, China)

机构地区：[1]中国水产科学研究院黄海水产研究所海洋产物资源与酶工程实验室,山东青岛266071

出　　处：《中国生物化学与分子生物学报》2018年第6期659-666,共8页Chinese Journal of Biochemistry and Molecular Biology

基　　金：中国水产科学研究院中央级公益性科研院所基本科研业务费专项资金(No.2017GH07&2017RC-YJ01);中国水产科学研究院黄海水产研究所中央级公益性科研院所基本科研业务费专项资金(No.20603022015012);国家自然科学基金(No.31402276)资助~~

摘　　要：蛋白酶MP(marine protease)是海洋细菌来源的新型碱性金属蛋白酶,在工业中具有良好的应用前景。本文采用酶动力学方法研究4-甲酰苯基硼酸(4-FPBA)对MP的抑制作用,并结合分子模拟的方法,通过水溶液环境下分子力学-玻尔兹曼泊松表面积(MM-PBSA)和量子力学/分子力学混合方法(QM/MM)对其抑制机制进行了研究。结果表明,4-FPBA对蛋白酶MP的抑制过程属可逆的竞争型抑制,抑制常数Ki为0.57 mmol/L。在4-FPBA与蛋白酶MP的结合过程中,范德瓦尔斯相互作用对于其结合发挥了重要作用。明确了Arg59、Leu151、His190和His196的4个残基为蛋白酶MP中与4-FPBA结合的关键残基。该研究结果将为今后进行蛋白酶MP可逆抑制剂的筛选与设计提供理论基础,以提高其液态稳定性,从而拓宽蛋白酶MP在液体洗涤剂中的高效应用。Marine protease（ MP） is a new alkaline metalloprotease derived from marine bacteria and has a good application prospect in industry. The inhibitory effect of 4-formyl-phenyl-boronic acid（ 4-FPBA）on MP was studied by enzyme kinetic methods and combined with molecular dynamics simulation. The inhibitory mechanism was studied by molecular mechanics/poisson Boltzmann surface area（ MM-PBSA）and quantum mechanics/molecular mechanics（ QM/MM） methods in explicit water. The results showed that the inhibition process of 4-FPBA on MP is reversible competitive inhibition and the inhibition constant Kiis 0. 57 mmol/L. Van Der Waals interaction plays an important role in the binding process of4-FPBA and MP. Furthermore,four residues Arg59,Leu151,His190 and His196 were identified as the key residues mediating the binding between MP and 4-FPBA. These results provide a theoretical basis for the further screening and rational design of MP inhibitors,which would improve its stability in the liquid state and also provide better applications in liquid detergents.

关 键 词：蛋白酶 可逆抑制 4-甲酰苯基硼酸 抑制机制 

分 类 号：Q556.9[生物学—生物化学] Q178.53