Lack of Evidence for a Role of Cys-138 as a Base Catalyst in the Skeletal Muscle 6-Phosphofructo-2-kinase Reaction

Irwin J. Kurland, M. R. Elmaghrabi, S. J. Pilkis

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)229-236
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume195
Issue number1
DOIs
StatePublished - Aug 31 1993
Externally publishedYes

Fingerprint

Phosphofructokinase-2
Muscle
Skeletal Muscle
Catalysts
Enzymes
Glutamine
Catalysis
Alanine
Serine
Escherichia coli
Catalytic Domain
Phosphotransferases
Kinetics

ASJC Scopus subject areas

  • Molecular Biology
  • Biophysics
  • Biochemistry

Cite this

Lack of Evidence for a Role of Cys-138 as a Base Catalyst in the Skeletal Muscle 6-Phosphofructo-2-kinase Reaction. / Kurland, Irwin J.; Elmaghrabi, M. R.; Pilkis, S. J.

In: Biochemical and Biophysical Research Communications, Vol. 195, No. 1, 31.08.1993, p. 229-236.

Research output: Contribution to journalArticle

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