Removal of substrate inhibition in a lactate dehydrogenase from human muscle by a single residue change

被引:56
作者
Eszes, CM
Sessions, RB
Clarke, AR
Moreton, KM
Holbrook, JJ
机构
[1] UNIV BRISTOL,SCH MED SCI,MOL RECOGNIT CTR,BRISTOL BS8 1TD,AVON,ENGLAND
[2] UNIV BRISTOL,SCH MED SCI,DEPT BIOCHEM,BRISTOL BS8 1TD,AVON,ENGLAND
关键词
lactate dehydrogenase; substrate inhibition; mutagenesis; enzyme kinetics; protein modelling;
D O I
10.1016/S0014-5793(96)01317-8
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
High concentrations of ketoacid substrates inhibit most natural hydroxyacid dehydrogenases due to the formation of an abortive enzyme-NAD(+)-ketoacid complex. It was postulated that this substrate inhibition could be eliminated from lactate dehydrogenases if the rate of NAD(+) dissociation could be increased, An analysis of the crystal structure of mammalian LDHs showed that the amide of the nicotinamide cofactor formed a water-bridged hydrogen bond to S163. The LDH of Plasmodium falciparum is not inhibited by its substrate and, uniquely, in this enzyme the serine is replaced by a leucine, In the S163L mutant of human LDH-M(4) pyruvate inhibition is, indeed, abolished and the enzyme retains high activity, However, the major contribution to this effect comes from a weakening of the interaction of pyruvate with the enzyme-coenzyme complex.
引用
收藏
页码:193 / 197
页数:5
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