Regulatory properties of purified 3-phosphoglycerate dehydrogenase from Bacillus subtilis.
3-Phosphoglycerate dehydrogenase (3-phosphoglycerate:NAD oxidoreductase, EC. 1.1.1.95) was purified from Bacillus subtilis by conventional methods. The final preparation was homogeneous by electrophoretic analysis and had a sedimentation constant of 6.3 S. On the basis of gel filtration data the enzyme had a molecular weight of about 166000. The plot of velocity versus phosphoglycerate concentration was biphasic while similar plots for hydroxypyruvate phosphate and NADH were the conventional hyperbolic type. The enzyme was specifically inhibited by serine. The inhibition was time dependent, requiring several minutes incubation before a constant level of inhibition was achieved. Serine inhibition was of the "mixed type" with respect to 3-phosphoglycerate and Hill plots of these data had slopes that approached 2. Desensitization of the enzyme to serine inhibition was achieved by incubation in the absence of dithiothreitol. The desensitized enzyme was different from the native enzyme in fluoresence properties, sedimentation characteristics and in the absence of the biphasic phosphoglycerate saturation curve. Evidence was obtained for the participation of sulphydryl groups in the changes in protein structure responsible for serine inhibition as well as the dehydrogenase activity of the enzyme.
D000429:Alcohol Oxidoreductases / Q000378:metabolism*; D001412:Bacillus subtilis / Q000201:enzymology*; D002237:Carbohydrate Dehydrogenases / Q000302:isolation & purification / Q000378:metabolism*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003460:Crystallization; D006902:Hydroxymercuribenzoates; D007700:Kinetics; D008970:Molecular Weight; D009243:NAD / Q000494:pharmacology; D010084:Oxidation-Reduction; D012694:Serine / Q000494:pharmacology; D013050:Spectrometry, Fluorescence; D013307:Streptomycin; D013997:Time Factors
D006902:Hydroxymercuribenzoates; D009243:NAD; D012694:Serine; D000429:Alcohol Oxidoreductases; D002237:Carbohydrate Dehydrogenases; D013307:Streptomycin