Site-directed mutagenesis of aspartic acid 372 at the ATP binding site of yeast phosphoglycerate kinase: over-expression and characterization of the mutant enzyme

In order to investigate the possible role of aspartate 372 in the kinase reaction, a mutant phosphoglycerate kinase (PGK) protein in which this residue is replaced by an asparagine, has been constructed by site-directed mutagenesis. This paper reports on the construction of this mutant PGK and its expression in yeast using a new yeast vector. It also reports on the kinetic properties of the resulting mutant enzyme relative to those of wild-type over-expressed PGK.