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Site-directed mutagenesis of histidine 62 in the 'basic patch' region of yeast phosphoglycerate kinase
W J Fairbrother1, L Hall, J A Littlechild
1Inorganic Chemistry Laboratory, Oxford, England.
FEBS Letters
|December 4, 1989
Abstract:
Site-directed mutagenesis has been used to produce a mutant form of yeast phosphoglycerate kinase (PGK) in which the 'basic patch' residue His 62 has been replaced by a glutamine residue. Using 1H-NMR spectroscopy, it was found that 3-phosphoglycerate (3-PG) binding to the mutant protein induces the same conformational effects as for wild-type PGK, although the affinity was reduced by 2- to 3-fold. Kinetic studies show both Km for 3-PG and Vmax to be increased by approximately 2-fold relative to the wild-type enzyme. These data are consistent with the suggestion that His 62 assists in the binding of the substrate to the enzyme.