Synthesis and properties of (4-13C)NAD+. Observation of its binding to yeast alcohol dehydrogenase by 13C-NMR spectroscopy.
Starting from (13C)formic acid, acetone and cyanoacetamide samples of (4-13C)nicotinic acid and (4-13C)-nicotinamide were synthesised in an overall and additive yield of 11%. 1H-NMR and mass spectroscopy showed 90% enrichment of 13C in the expected position. NADase-catalysed exchange between thionicotinamide-adenine dinucleotide and (4-13C)nicotinamide furnished (4-13C)NAD+ which was purified, characterized and quantified by 1H-NMR and 13C-NMR spectroscopy and by enzymic assay. The 13C-NMR signal of (4-13C)beta-NAD+ (146.09 ppm) was broadened and shifted (147.83 ppm) upon binding to yeast alcohol dehydrogenase.
D000426:Alcohol Dehydrogenase; D000429:Alcohol Oxidoreductases / Q000378:metabolism*; D001665:Binding Sites; D002247:Carbon Isotopes; D002384:Catalysis; D055598:Chemical Phenomena; D002621:Chemistry; D007553:Isotope Labeling / Q000379:methods; D009682:Magnetic Resonance Spectroscopy; D009243:NAD / Q000138:chemical synthesis / Q000378:metabolism*; D009525:Niacin / Q000138:chemical synthesis; D009536:Niacinamide / Q000138:chemical synthesis; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000201:enzymology*
D002247:Carbon Isotopes; D009243:NAD; D009536:Niacinamide; D009525:Niacin; D000429:Alcohol Oxidoreductases; D000426:Alcohol Dehydrogenase