Arg-94 is crucial to the catalysis of 3-isopropylmalate dehydrogenase from Thermus thermophilus HB8

被引：4

作者：

Fujita, M ^{[1
]}

Toyooka, Y ^{[1
]}

Tamegai, H ^{[1
]}

Eguchi, T ^{[1
]}

Kakinuma, K ^{[1
]}

机构：

[1] Tokyo Inst Technol, Dept Chem, Meguro Ku, Tokyo 1528551, Japan

来源：

JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC | 2000年 / 9卷 / 4-6期

关键词：

Thermus thermophilus HB-8; 3-isopropylmalate dehydrogenase; site-directed mutagenesis; catalytic residue;

D O I：

10.1016/S1381-1177(99)00091-0

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 crucial role of Arg-94 in 3-isopropylmalate (IPM) dehydrogenase from Thermus thermophilus HB8 was elucidated by replacing the residue to lysine with site-directed mutagenesis. The k(cat) value of the R94K mutant enzyme for IPM was significantly reduced to 1/170 compared with that of native enzyme, whereas the K-m for IPM was not much changed. It appeared that the major role of Arg-94 in exerting the enzymatic activity is not for the substrate recognition, but for the reaction catalysis, in such a way that Arg-94 facilitates stabilization of the transition-state in the decarboxylation step. (C) 2000 Elsevier Science B.V. All rights reserved.

引用

页码：149 / 155

页数：7

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