title: On the effect on specificity of Thr246----Gly mutation in L-lactate dehydrogenase of Bacillus sterothermophilus.
abstract: The function of the amino acid Thr246 in L-lactate dehydrogenase from Bacillus stearothermophilus has been investigated by site-directed replacement with glycine. Kinetic experiments with a number of 2-oxo acids showed strongly reduced activity for the mutated enzyme. However, the mutant enzyme shows a relative preference for the large hydrophobic sidechains of alpha-keto acids and an even higher specific activity than the wild-type lactate dehydrogenase for the polar oxaloacetate substrate. Graphic analyses indicate that the loss of one hydrogen bond, or intrusion of water into the active site, might be responsible for the reduced activity. The kinetic results suggest that the binding modes of bulky hydrophobic or polar substrates compensate to some degree for the partially disrupted active site.
authors.affiliation: Department of Chemistry, University of Toronto, Ontario, Canada.
authors.forename: D
authors.initials: D
authors.lastname: Bur
authors.forename: T
authors.initials: T
authors.lastname: Clarke
authors.forename: J D
authors.initials: JD
authors.lastname: Friesen
authors.forename: M
authors.initials: M
authors.lastname: Gold
authors.forename: K W
authors.initials: KW
authors.lastname: Hart
authors.forename: J J
authors.initials: JJ
authors.lastname: Holbrook
authors.forename: J B
authors.initials: JB
authors.lastname: Jones
authors.forename: M A
authors.initials: MA
authors.lastname: Luyten
authors.forename: H M
authors.initials: HM
authors.lastname: Wilks
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002384:Catalysis; D005798:Genes, Bacterial; D001411:Geobacillus stearothermophilus / Q000201:enzymology* / Q000235:genetics; D005998:Glycine / Q000235:genetics*; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000235:genetics*; D009154:Mutation; D013379:Substrate Specificity; D013912:Threonine / Q000235:genetics*
chemical_list: D013912:Threonine; D007770:L-Lactate Dehydrogenase; D005998:Glycine