ROLE OF THR-252 IN CYTOCHROME P450(CAM) - A STUDY WITH UNNATURAL AMINO-ACID MUTAGENESIS

被引：108

作者：

KIMATA, Y

SHIMADA, H

HIROSE, T

ISHIMURA, Y

机构：

[1] KEIO UNIV, SCH MED, DEPT BIOCHEM, SHINJUKU KU, TOKYO 160, JAPAN

[2] KEIO UNIV, SCH MED, INST PHARMACEUT, SHINJUKU KU, TOKYO 160, JAPAN

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 1995年 / 208卷 / 01期

关键词：

D O I：

10.1006/bbrc.1995.1310

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：

Replacement of Thr-252 in the active center of cytochrome P450cam with a non-hydroxy amino acid residue such as Ala and Val by conventional site-directed mutagenesis converted this monooxygenase to an NADH oxidase (Imai, M. et al. Proc. Natl. Sci. U. S. A. 86, 7823-7827, 1989). In this study, a mutant enzyme with a methoxy group in place of the hydroxy group of Thr-252 (OMe-mutant) was synthesized by the method of unnatural amino acid mutagenesis (Noren, C. J. et al., Science 244, 182-188, 1989). Unlike other site-directed mutants without a hydroxy group at the position, the OMe-mutant retained a considerably high monooxygenase activity, yielding a stoichiometric amount of 5-exo-hydroxycamphor to that of the oxygen consumed. Thus a free hydroxy group at this position is not an indispensable requisite for the monooxygenase to cleave the O-O bond of molecular O-2 as previously proposed. (C) 1995 Academic Press. Inc.

引用

页码：96 / 102

页数：7

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