EXPRESSION AND SITE-DIRECTED MUTAGENESIS OF HEPATIC GLUCOKINASE

被引：32

作者：

LANGE, AJ ^{[1
]}

XU, LZ ^{[1
]}

VANPOELWIJK, F ^{[1
]}

LIN, K ^{[1
]}

GRANNER, DK ^{[1
]}

PILKIS, SJ ^{[1
]}

机构：

[1] VANDERBILT UNIV, MED CTR, SCH MED, DEPT MOLEC PHYSIOL & BIOPHYS, NASHVILLE, TN 37232 USA

来源：

BIOCHEMICAL JOURNAL | 1991年 / 277卷

关键词：

D O I：

10.1042/bj2770159

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：

Soluble rat liver glucokinase was expressed at high levels at 22-degrees-C in the BL21(DE3)pLysS strain of Escherichia coli. Aspartate-211 of yeast hexokinase has been implicated as a catalytic residue from crystallographic data. The corresponding residue in rat liver glucokinase, aspartate-205, was mutated to alanine and the expressed mutant had 1/500th of the activity of the wild type, with no change in the K(m) values for glucose or ATP. The results support a role for this residue as a base catalyst in the glucokinase reaction and, most probably, a similar role in the reactions of all members of the hexokinase family.

引用

页码：159 / 163

页数：5

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