Site‐directed mutagenesis of AMP‐binding residues in adenylate kinase Alteration of substrate specificity

FEBS Letters

Okajima, Toshihide¹  Fukui, Toshio¹  Tanizawa, Katsuyuki¹ 

[1] Institute of Scientific and Industrial Research, Osaka University, Ibaraki, Osaka 567, Japan

关键词: Adenylate kinase;    UMP-CMP kinase;    Site-directed mutagenesis;    Substrate specificity;    AMP-binding;    PCR;    polymerase chain reaction;    T39A and L66I;    the single mutant enzymes in which Thr39 and Leu66 are separately replaced by Ala and Ile;    respectively;    T39A/L66I;    the double mutant enzyme in which Thr39 and Leu66 are concomitantly replaced by Ala and Ile;    respectively;   

DOI  :  10.1016/0014-5793(93)81687-U

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Adenylate kinase is highly specific for AMP as phosphoryl acceptor. We have found that the replacement of Thr³⁹ by Ala in the chicken muscle enzyme, alone or together with the replacement of Leu⁶⁶ by Ile, caused remarkable increases in CMP and UMP activities with a concomitant decrease in AMP activity; therefore, the resulting mutant enzymes show CMP and UMP activities/AMP activity ratios much higher than the wild-type enzyme. The mutant enzyme in which Ala is substituted for Thr³⁹ has a V _max value for CMP comparable to that of CMP-UMP kinase.

【 授权许可】

Unknown