1-脱氧-D-木酮糖 5-磷酸合酶催化一种新颖的随机顺序机制。
1-Deoxy-D-xylulose 5-phosphate synthase catalyzes a novel random sequential mechanism.
机构信息
Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
出版信息
J Biol Chem. 2011 Oct 21;286(42):36522-31. doi: 10.1074/jbc.M111.259747. Epub 2011 Aug 30.
Abstract
Emerging resistance of human pathogens to anti-infective agents make it necessary to develop new agents to treat infection. The methylerythritol phosphate pathway has been identified as an anti-infective target, as this essential isoprenoid biosynthetic pathway is widespread in human pathogens but absent in humans. The first enzyme of the pathway, 1-deoxy-D-xylulose 5-phosphate (DXP) synthase, catalyzes the formation of DXP via condensation of D-glyceraldehyde 3-phosphate (D-GAP) and pyruvate in a thiamine diphosphate-dependent manner. Structural analysis has revealed a unique domain arrangement suggesting opportunities for the selective targeting of DXP synthase; however, reports on the kinetic mechanism are conflicting. Here, we present the results of tryptophan fluorescence binding and kinetic analyses of DXP synthase and propose a new model for substrate binding and mechanism. Our results are consistent with a random sequential kinetic mechanism, which is unprecedented in this enzyme class.
摘要
人类病原体对抗感染药物的耐药性不断出现，使得开发新的药物来治疗感染变得非常必要。甲羟戊酸途径已被确定为一种抗感染的靶标，因为这条必需的异戊烯醇生物合成途径在人类病原体中广泛存在，但在人类中不存在。该途径的第一个酶，1-脱氧-D-木酮糖 5-磷酸（DXP）合酶，通过在硫胺素二磷酸依赖性方式下缩合 D-甘油醛 3-磷酸（D-GAP）和丙酮酸来催化 DXP 的形成。结构分析揭示了一种独特的结构域排列，这为 DXP 合酶的选择性靶向提供了机会；然而，关于其动力学机制的报告却存在矛盾。在这里，我们介绍了 DXP 合酶的色氨酸荧光结合和动力学分析的结果，并提出了一个新的底物结合和机制模型。我们的结果与随机顺序动力学机制一致，这在该酶类中是前所未有的。
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