Significance of the enzymatic properties of yeast S39A enolase to the catalytic mechanism

被引:20
作者
Brewer, JM [1 ]
Glover, CVC
Holland, MJ
Lebioda, L
机构
[1] Univ Georgia, Dept Biochem & Mol Biol, Athens, GA 30602 USA
[2] Univ Calif Davis, Dept Biol Chem, Sch Med, Davis, CA 95616 USA
[3] Univ S Carolina, Dept Chem & Biochem, Columbia, SC 29208 USA
来源
BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY | 1998年 / 1383卷 / 02期
关键词
enolase; mutagenesis; catalytic mechanism; loop closure;
D O I
10.1016/S0167-4838(98)00004-1
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 S39A mutant of yeast enolase (isozyme 1), prepared by site-directed mutagenesis, has a relative V-max of 0.01% and an activation constant for Mg2+ ca. 10-fold higher, compared with native enzyme. It is correctly folded. There is little effect of solvent viscosity on activity. We think that the loop Ser36-His43 fails to move to the 'closed' position upon catalytic Mg2+ binding, weakening several electrostatic interactions involved in the mechanism. (C) 1998 Elsevier Science B.V.
引用
收藏
页码:351 / 355
页数:5
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