羧肽酶A的作用机制：酮底物类似物的水合作用
Mechanism of carboxypeptidase A: hydration of a ketonic substrate analogue.
作者信息
Christianson D W, David P R, Lipscomb W N
出版信息
Proc Natl Acad Sci U S A. 1987 Mar;84(6):1512-5. doi: 10.1073/pnas.84.6.1512.
Abstract
The structure of the complex between carboxypeptidase A alpha (EC 3.4.17.1) and the ketonic substrate analogue 5-benzamido-2-benzyl-4-oxopentanoic acid (BOP) has been determined by x-ray crystallographic methods to a resolution of 1.7 A (final R = 0.191). Interestingly, BOP was observed to bind to the active site of carboxypeptidase A alpha as the covalent hydrate adduct. Because BOP is probably less than 0.2% hydrated in aqueous solution, this result was unexpected. One possibility is that the zinc-bound water of the native enzyme added to the ketone carbonyl. Alternatively, the enzyme may preferentially scavenge the hydrated ketone as it is continuously maintained at equilibrium in the solution in which the carboxypeptidase A alpha crystals were immersed. In either case, this mode of binding of BOP to carboxypeptidase A alpha provides an example of the preferred binding of a model of a structure along the reaction coordinate of a hydrolytic reaction.
摘要
通过X射线晶体学方法确定了羧肽酶Aα（EC 3.4.17.1）与酮底物类似物5-苯甲酰胺基-2-苄基-4-氧代戊酸（BOP）之间复合物的结构，分辨率为1.7埃（最终R = 0.191）。有趣的是，观察到BOP以共价水合物加合物的形式结合到羧肽酶Aα的活性位点。由于BOP在水溶液中的水合程度可能小于0.2%，这个结果出乎意料。一种可能性是天然酶中与锌结合的水加到了酮羰基上。或者，在羧肽酶Aα晶体浸入的溶液中，该酶可能优先清除处于平衡状态的水合酮。无论哪种情况，BOP与羧肽酶Aα的这种结合模式提供了一个沿着水解反应坐标的结构模型优先结合的例子。
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