Lack of Evidence for a Role of Cys-138 as a Base Catalyst in the Skeletal Muscle 6-Phosphofructo-2-kinase Reaction
| Autor: | M R el-Maghrabi, S J Pilkis, Irwin J. Kurland |
|---|---|
| Rok vydání: | 1993 |
| Předmět: |
Phosphofructokinase-1
Molecular Sequence Data Biophysics Polymerase Chain Reaction Biochemistry Serine Escherichia coli medicine Animals Point Mutation T7 RNA polymerase Amino Acid Sequence Cysteine Cloning Molecular Site-directed mutagenesis Molecular Biology Alanine chemistry.chemical_classification Base Sequence biology Muscles Active site Skeletal muscle Cell Biology Recombinant Proteins Glutamine Kinetics Enzyme medicine.anatomical_structure Oligodeoxyribonucleotides chemistry Mutagenesis Site-Directed biology.protein medicine.drug |
| Zdroj: | Biochemical and Biophysical Research Communications. 195:229-236 |
| ISSN: | 0006-291X |
| DOI: | 10.1006/bbrc.1993.2034 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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