THE COOPERATIVITY AND ALLOSTERIC INHIBITION OF ESCHERICHIA-COLI PHOSPHOFRUCTOKINASE DEPEND ON THE INTERACTION BETWEEN THREONINE-125 AND ATP

被引:17
作者
AUZAT, I [1 ]
LEBRAS, G [1 ]
GAREL, JR [1 ]
机构
[1] CNRS,ENZYMOL LAB,F-91198 GIF SUR YVETTE,FRANCE
关键词
PHOSPHORYL TRANSFER; SITE DIRECTED MUTAGENESIS; ENZYME REGULATION;
D O I
10.1073/pnas.91.12.5242
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
During the reaction catalyzed by the phosphofructokinase (EC 2.7.1;11) from Escherichia coli, the phosphoryl group transferred from ATP interacts with Thr-125 [Shirakihara, Y. and Evans, P. R. (1988) J. Mel. Biol. 204, 973-995]. The replacement of Thr-125 by serine changes the saturation by fructose 6-phosphate from cooperative to hyperbolic and abolishes the allosteric inhibition by phosphoenolpyruvate. The same changes, a saturation by fructose 6-phosphate that is no longer cooperative and an activity that is no longer inhibited by phosphoenolpyruvate, are observed with wild-type phosphofructokinase when adenosine 5'-[gamma-thio]triphosphate is used instead of ATP as the phosphoryl donor. These two perturbations of the ATP-Thr-125 interaction lead to the suppression of both the allosteric inhibition by phosphoenolpyruvate and the cooperativity of fructose-6-phosphate saturation, as if replacing the neutral oxygen of ATP by sulfur or removing the methyl group of Thr-125 had ''locked'' phosphofructokinase in its active conformation. The geometry of this ATP-Thr-125 interaction and/or the presence of the methyl group on the beta-carbon of Thr-125 are crucial for the regulatory properties of phosphofructokinase. This interaction could be a hydrogen bond between the neutral oxygen of the gamma-phosphate of ATP and the hydroxyl group of Thr-125.
引用
收藏
页码:5242 / 5246
页数:5
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