title: Mitochondrial alpha-glycerol phosphate dehydrogenase activity in IIA fibres of the rat lateral gastrocnemius muscle; the effect of Ca2+ and ATP.
abstract: Mitochondrial alpha-glycerol phosphate dehydrogenase is an important enzyme, but it is difficult to extract and purify. We have measured the activity of this enzyme in single type IIA skeletal muscle fibres under initial rate conditions by microdensitometry of the formazan reaction product. The Km (1.6 mM) for the substrate (L-alpha-glycerol phosphate) was lower than reported for the extracted enzyme. Further, at low substrate concentrations (3 mM), the enzyme was allosterically activated by free Ca2+ concentrations of 1 microM or greater, and half-maximal stimulation occurred at 0.3 microM free Ca2+. In the absence of Ca2+, there was negative cooperativity of substrate binding with a Hill constant of 0.57, but no cooperativity occurred in the presence of calcium. ATP (10 mM) inhibited enzyme activity in the presence of Ca2+ but not in its absence.
authors.affiliation: Department of Medicine (Clinical Biochemistry), University of Manchester, Salford, UK.
authors.forename: R B
authors.initials: RB
authors.lastname: Lomax
authors.forename: W R
authors.initials: WR
authors.lastname: Robertson
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D003720:Densitometry / Q000379:methods; D004305:Dose-Response Relationship, Drug; D005562:Formazans / Q000378:metabolism; D005993:Glycerolphosphate Dehydrogenase / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D008931:Mitochondria, Muscle / Q000201:enzymology*; D009132:Muscles / Q000648:ultrastructure*; D051381:Rats; D011919:Rats, Inbred Strains
chemical_list: D005562:Formazans; D000255:Adenosine Triphosphate; D005993:Glycerolphosphate Dehydrogenase; D002118:Calcium