The mechanism of the alkaline phosphatase reaction: insights from NMR, crystallography and site-specific mutagenesis

被引：135

作者：

Holtz, KM ^{[1
]}

Kantrowitz, ER ^{[1
]}

机构：

[1] Boston Coll, Dept Chem, Merkert Chem Ctr, Chestnut Hill, MA 02467 USA

来源：

FEBS LETTERS | 1999年 / 462卷 / 1-2期

关键词：

mechanism; phosphatase; rate-limiting step; site-specific mutagenesis; transphosphorylation; trigonal bipyramidal;

D O I：

10.1016/S0014-5793(99)01448-9

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The proposed double in-line displacement mechanism of Escherichia coli alkaline phosphatase (AP) involving two-metal ion catalysis is based on NMR spectroscopic and X-ray crystallographic studies. This mechanism is further supported by the X-ray crystal structures of the covalent phospho-enzyme intermediate of the H331Q mutant AP and of the transition state complex between the mild-type enzyme and vanadate, a transition state analog, Kinetic and structural studies on several genetically engineered versions of AP illustrate the overall importance of the active site's metal geometry, hydrogen bonding network and electrostatic potential in the catalytic mechanism, (C) 1999 Federation of European Biochemical Societies.

引用

页码：7 / 11

页数：5

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