2-oxoglutarate dehydrogenase complex from bovine-adrenal-cortex mitochondria. Purification and partial characterization.
The 2-oxoglutarate dehydrogenase complex from bovine adrenal-cortex mitochondria has been purified by polyethylene glycol fractionation, ultracentrifugation through a layer of sucrose, isoelectric precipitation and gel filtration of Sepharose 4 B. The specific activity of the preparation obtained wa 9.9 U/mtg of protein; the sedimentation coefficient, S20, w, was 30 S. The results of sodium dodecyl sulphate polyacrylamide gel electrophoresis indicated decomposition of the 2-oxoglutarate dehydrogenase complex into 3 clear-cut protein fractions with mobilities corresponding to molecular weights of 51 000, 56 000 and about 110 000. Michaelis constants for the reactants of the 2-oxoglutarate dehydrogenase complex reactions were: 2-oxoglutarate = 200 micro M; CoA - 4,5 micro M; NAD - 25 micro M.
D000302:Adrenal Cortex / Q000201:enzymology*; D000818:Animals; D002417:Cattle; D002499:Centrifugation, Density Gradient; D011232:Chemical Precipitation; D002850:Chromatography, Gel; D003065:Coenzyme A / Q000378:metabolism; D007655:Ketoglutarate Dehydrogenase Complex / Q000302:isolation & purification / Q000378:metabolism*; D007656:Ketoglutaric Acids / Q000378:metabolism; D007658:Ketone Oxidoreductases / Q000378:metabolism*; D007700:Kinetics; D008928:Mitochondria / Q000201:enzymology*; D009243:NAD / Q000378:metabolism; D011092:Polyethylene Glycols
D007656:Ketoglutaric Acids; D009243:NAD; D011092:Polyethylene Glycols; D007658:Ketone Oxidoreductases; D007655:Ketoglutarate Dehydrogenase Complex; D003065:Coenzyme A