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

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.