A simple method for the rapid determination of the stereospecificity of NAD-dependent dehydrogenases applied to mammalian IMP dehydrogenase and bacterial NADH peroxidase.
The stereospecificity of IMP dehydrogenase (IMP:NAD+ oxidoreductase, EC 1.1.1.205) from two different sources was determined. The enzyme preparations were obtained from murine lymphoblasts and from Escherichia coli. Both enzymes transferred the 2-3H of IMP to the pro-S position of carbon atom C-4 of the nicotinamide ring in NAD. Thus, B-sided stereospecificity is common to the enzyme from two very different species. In addition, the studies described here demonstrate that alcohol dehydrogenase and NADH peroxidase, used as auxiliary enzymes, in combination with a microdistillation procedure, should permit rapid determination of the stereospecificity of any NAD-dependent dehydrogenase for which the appropriate tritiated substrate is available.
D000426:Alcohol Dehydrogenase / Q000378:metabolism; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004754:Enterobacter / Q000201:enzymology; D004926:Escherichia coli / Q000201:enzymology; D007168:IMP Dehydrogenase / Q000378:metabolism*; D007658:Ketone Oxidoreductases / Q000378:metabolism*; D051379:Mice; D010544:Peroxidases / Q000378:metabolism*; D013237:Stereoisomerism; D013379:Substrate Specificity
D000426:Alcohol Dehydrogenase; D007168:IMP Dehydrogenase; D010544:Peroxidases; C021845:NAD+ peroxidase; D007658:Ketone Oxidoreductases