Biochemical analyses of natural and induced null variants of Drosophila enzymes.
For the characterization of null mutants identified in Drosophila populations, several Drosophila enzymes including alcohol dehydrogenase, cytoplasmic malate dehydrogenase, alpha-glycerol phosphate dehydrogenase, and phosphoglucose isomerase were co-purified to homogeneity using an 8-(6-aminohexyl)-amino-ATP-Sepharose affinity column followed by DEAE-Sepharose column chromatography. Mitochondrial malate dehydrogenase was purified by the use of CM-Sepharose and the same ATP affinity column. Alcohol dehydrogenase and alpha-glycerol phosphate dehydrogenase were mapped on a two-dimensional gel. Antiserum was raised in rabbits against these Drosophila enzymes. The presence of cross-reacting material in null mutants was characterized by double immunodiffusion, immunoelectrophoresis, and two-dimensional gel electrophoresis. By immunological techniques, two natural null variants of malic enzyme and one of phosphoglucose isomerase were shown to be negative to cross-reacting material. Two low-dose-rate gamma-radiation-induced null mutants of cytoplasmic malate dehydrogenase were shown to be positive to cross-reacting material. Two-dimensional gel analyses enabled the characterization of three natural null variants of alpha-glycerol phosphate dehydrogenase. The viability of some null mutants with homozygous null or null/deficiency genotypes is discussed in terms of the in vivo metabolic roles of the related enzymes.
D000429:Alcohol Oxidoreductases / Q000235:genetics* / Q000302:isolation & purification; D000818:Animals; D004331:Drosophila melanogaster / Q000201:enzymology* / Q000235:genetics; D014644:Genetic Variation; D005956:Glucose-6-Phosphate Isomerase / Q000235:genetics* / Q000302:isolation & purification; D005993:Glycerolphosphate Dehydrogenase / Q000235:genetics* / Q000302:isolation & purification; D005779:Immunodiffusion; D007122:Immunoelectrophoresis; D008291:Malate Dehydrogenase / Q000235:genetics* / Q000302:isolation & purification; D009154:Mutation
D000429:Alcohol Oxidoreductases; D005993:Glycerolphosphate Dehydrogenase; D008291:Malate Dehydrogenase; D005956:Glucose-6-Phosphate Isomerase