Affinity precipitation of dehydrogenases.
Affinity precipitation, a novel technique closely related to immunoprecipitation and affinity chromatography, has been evaluated in systems comprised of dehydrogenases and a bifunctional NAD derivative, Bis-NAD. Lactate dehydrogenase and glutamate dehydrogenase were easily precipitated whereas yeast alcohol dehydrogenase required the presence of salt to enhance the affinity precipitation. Liver alcohol dehydrogenase did not precipitate, probably because most of the affinity complexes formed were composed of only two enzyme molecules. Affinity precipitation was carried out on a preparative scale for the isolation of ox heart lactate dehydrogenase from a crude extract. The yield and purity of the enzyme and the general properties of the procedure are considered very satisfactory.
D000345:Affinity Labels; D000426:Alcohol Dehydrogenase; D000429:Alcohol Oxidoreductases / Q000302:isolation & purification; D000818:Animals; D002417:Cattle; D011232:Chemical Precipitation; D005969:Glutamate Dehydrogenase / Q000302:isolation & purification; D007770:L-Lactate Dehydrogenase / Q000302:isolation & purification; D008099:Liver / Q000201:enzymology; D009206:Myocardium / Q000201:enzymology; D009243:NAD / Q000031:analogs & derivatives; D010088:Oxidoreductases / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae / Q000201:enzymology
D000345:Affinity Labels; D009243:NAD; C018812:N(2),N(2)'-adipodihydrazido-bis-(N(6)-carbonylmethyl-NAD); D010088:Oxidoreductases; D000429:Alcohol Oxidoreductases; D000426:Alcohol Dehydrogenase; D007770:L-Lactate Dehydrogenase; D005969:Glutamate Dehydrogenase