Article
Site-directed mutagenesis of aspartic acid 372 at the ATP binding site of yeast phosphoglycerate kinase: over-expression and characterization of the mutant enzyme.
Protein engineering - 1 May 1990
Minard P, Bowen D J, Hall L, Littlechild J A, Watson H C
Abstract excerpt
A new phosphoglycerate kinase over-expression vector, pYE-PGK, has been constructed which greatly facilitates the insertion and removal of mutant enzyme genes by cleavage at newly introduced BamHI sites. This vector has been used to prepare mutant protein in appreciable (100 mg) quantities for use in kinetic, crystallographic and NMR experiments. Aspartate 372 is an invariant amino acid residue in genes known to...
Topics
- Adenosine Triphosphate
- Amino Acid Sequence
- Aspartic Acid
- Base Sequence
- Binding Sites
- Escherichia coli
- Gene Expression
- Genetic Vectors
- Kinetics
- Molecular Sequence Data
- Mutation