MODIFICATION OF THE ACTIVE-SITE OF ALKALINE-PHOSPHATASE BY SITE-DIRECTED MUTAGENESIS

被引：53

作者：

GHOSH, SS ^{[1
]}

BOCK, SC ^{[1
]}

ROKITA, SE ^{[1
]}

KAISER, ET ^{[1
]}

机构：

[1] ROCKEFELLER UNIV, BIOORGAN CHEM & BIOCHEM LAB, NEW YORK, NY 10021 USA

来源：

SCIENCE | 1986年 / 231卷 / 4734期

关键词：

D O I：

10.1126/science.3510454

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

The catalytically essential amino acid in the active site of bacterial alkaline phosphatase (Ser-102) has been replaced with a cysteine by site-directed mutagenesis. The resulting thiol enzyme catalyzes the hydrolysis of a variety of phosphate monoesters. The rate-determining step of hydrolysis, however, is no longer the same for catalysis when the active protein nucleophile is changed from the hydroxyl of serine to the thiol of cysteine. Unlike the steady-state kinetics of native alkaline phosphatase, those of the mutant show sensitivity to the leaving group of the phosphate ester.

引用

页码：145 / 148

页数：4

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