TY - JOUR
T1 - Lack of evidence for a role of Cys-138 as a base catalyst in the skeletal muscle 6-phosphofructo-2-kinase reaction
AU - Kurland, Irwin J.
AU - Elmaghrabi, M. Raafar
AU - Pilkis, Simon J.
N1 - Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.
PY - 1993/8/31
Y1 - 1993/8/31
N2 - The role of Cys-138 in the catalysis of the skeletal muscle 6-phosphofructo-2-kinase reaction was investigated by mutating this residue to serine, glutamine and alanine, expressing the mutants in E. coli with a T7 RNA polymerase-based expression system, and analyzing their kinetic properties. The Cys138Ala mutant had greatly diminished activity, while the Cys138Ser and Cys138Gln mutants had maximal velocities 2-3 fold higher than the wild-type enzyme. It was concluded that Cys-138 does not act as a base catalyst in the kinase reaction, but that it plays a significant structural role in the enzyme′s active site.
AB - The role of Cys-138 in the catalysis of the skeletal muscle 6-phosphofructo-2-kinase reaction was investigated by mutating this residue to serine, glutamine and alanine, expressing the mutants in E. coli with a T7 RNA polymerase-based expression system, and analyzing their kinetic properties. The Cys138Ala mutant had greatly diminished activity, while the Cys138Ser and Cys138Gln mutants had maximal velocities 2-3 fold higher than the wild-type enzyme. It was concluded that Cys-138 does not act as a base catalyst in the kinase reaction, but that it plays a significant structural role in the enzyme′s active site.
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U2 - 10.1006/bbrc.1993.2034
DO - 10.1006/bbrc.1993.2034
M3 - Article
C2 - 8363605
AN - SCOPUS:0027320571
SN - 0006-291X
VL - 195
SP - 229
EP - 236
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -