Investigations on myo-inositol-1-phosphate synthase from the wild type and the inositol-dependent mutant of Neurospora crassa.
The inositol-dependent mutant of Neurospora crassa lacks inositol-1-phosphate synthetase activity. This defect can be revorted by the addition of high-molecular DNA isolated from the wild type. To elucidate the biochemical background of inositol dependence, inositol-1-phosphate synthetase was studied. A method has been developed fro the isolation of the enzyme from the wild type strain in 10 mg scale by salt fractionation, gel filtration and ion-exchange chromatography. The specific activity of the purified enzyme is 4750 U/mg protein and its purity has increased about 100-fold. Polyacrylamide gel electrophoresis indicated that, in addition to the main enzymatically active band, several accompanying proteins occur in very small amount. The molecular weight of the enzyme is 225,000 daltons. Probably it consists of four subunits, two with a molecular weight of 64,000 daltons and another two of 50,000 daltons. An enzymatically inactive protein has been isolated from the mutant with the same procedure as that of the enzyme; it migrated at gel electrophoreis similarly to the enzyme. It may be supposed that the isolated protein is the defective enzyme molecule.
D002238:Carbohydrate Epimerases / Q000378:metabolism*; D007294:Inositol / Q000378:metabolism*; D046911:Macromolecular Substances; D008970:Molecular Weight; D009154:Mutation; D007296:Myo-Inositol-1-Phosphate Synthase / Q000302:isolation & purification / Q000378:metabolism*; D009491:Neurospora / Q000201:enzymology*; D009492:Neurospora crassa / Q000201:enzymology* / Q000235:genetics
D046911:Macromolecular Substances; D007294:Inositol; D002238:Carbohydrate Epimerases; D007296:Myo-Inositol-1-Phosphate Synthase