Characterization of the transition-state structure of the reaction of kanamycin nucleotidyltransferase by heavy-atom kinetic isotope effects

被引：19

作者：

Gerratana, B ^{[1
]}

Frey, PA ^{[1
]}

Cleland, WW ^{[1
]}

机构：

[1] Univ Wisconsin, Coll Agr & Life Sci, Dept Biochem, Inst Enzyme Res, Madison, WI 53705 USA

来源：

BIOCHEMISTRY | 2001年 / 40卷 / 09期

关键词：

D O I：

10.1021/bi002557x

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 transition-state structure for the reaction catalyzed by kanamycin nucleotidyltransferase has been determined from kinetic isotope effects. The primary (18)O isotope effects at pH 5.7 (close to the optimum pH) and at pH 7.7 (away from the optimum pH) are respectively 1.016 +/- 0.003 and 1.014 +/- 0.002. Secondary (18)O isotope effects of 1.0033 +/- 0.0004 and 1.0024 +/- 0.0002 for both nonbridge oxygen atoms were measured respectively at pH 5.7 and 7.7. These isotope effects are consistent with a concerted reaction with a slightly associative transition-state structure.

引用

页码：2972 / 2977

页数：6

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