迈向酶的计算机辅助定点诱变
Toward computer-aided site-directed mutagenesis of enzymes.
作者信息
Warshel A, Sussman F
出版信息
Proc Natl Acad Sci U S A. 1986 Jun;83(11):3806-10. doi: 10.1073/pnas.83.11.3806.
Abstract
A preliminary attempt to simulate the observed effect of a site-directed mutagenesis of rat trypsin gives encouraging results. The calculations reproduce in a semiquantitative way the observed change in the activation barrier of the rate-limiting step of amide hydrolysis. This result, which did not require any adjustable parameters, indicates that our method may provide a reliable basis for computer-aided enzyme design. In addition to the potentially practical value of the calculations, they provide important mechanistic information--that is, the change in the catalytic effect in trypsin appears to be almost exclusively due to the change in the electrostatic stabilization of the ionic configurations. This supports the view that electrostatic effects are the major factor in enzyme catalysis.
摘要
对大鼠胰蛋白酶定点诱变所观察到的效应进行初步模拟，得到了令人鼓舞的结果。计算结果以半定量的方式重现了酰胺水解限速步骤活化能垒中观察到的变化。这一结果无需任何可调参数，表明我们的方法可为计算机辅助酶设计提供可靠依据。除了这些计算具有潜在的实用价值外，它们还提供了重要的机理信息，即胰蛋白酶催化效应的变化似乎几乎完全归因于离子构型静电稳定性的变化。这支持了静电效应是酶催化主要因素的观点。
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