Purification and biochemical properties of allelic forms of cytoplasmic glycerol-3-phosphate dehydrogenase from Drosophila virilis.

Three homozygous allelic forms (alpha GPDHf, alpha GPDHm and alpha GPDHs) of NAD+-dependent glycerol-3-phosphate dehydrogenase (sn-glycerol-3-phosphate:NAD+ 2-oxidoreductase, EC 1.1.1.8) of Drosophila virilis were purified to homogeneity and their biochemical properties were compared. Although these three forms were mutually distinguishable by electrophoresis, no significant differences were found with respect to pH optima for both forward and reverse reactions (pH 6.0--6.5 for dihydroxyacetone phosphate reduction; pH 10.0--10.5 for glycerol 3-phosphate oxidation), native and subunit molecular weights (65 000 for native form; 35 000--37 000 for subunit) and Michaelis constants for NADH, glycerol 3-phosphate and NAD+ (5.3--6.0 microM for NADH; 1.8--1.9 mM for glycerol 3-phosphate; 100--110 microM for NAD+). Significant differences among three forms were observed in thermostability at 35 degrees C and inhibition by excess of dihydroxyacetone phosphate. The alpha GPDHf form was found to be most thermolabile and the alpha GPDHs form most susceptible to the inhibition.