Purification and properties of 6-phosphogluconate dehydrogenase from rabbit mammary gland.
Biochem J, 1975/11;151(2):263-70.
PMID: 3163
Impact factor: 3.766
Abstract
1. 6-Phosphogluconate dehydrogenase from rabbit mammary gland was purified to homogeneity by the criterion of polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate. The molecular weight of the subunit is 52 000. The enzyme was purified 150-fold with a final specific activity of 20 mumol of NADP+ reduced/min per mg of protein and overall yield of 3%. The molecular weight of the native enzyme is estimated to be 104 000 from gel-filtration studies. The final purification step was carried out by affinity chromatography with NADP+-Sepharose. 2. The Km values for 6-phosphogluconate and NADP+ are approx. 54 muM and 23 muM respectively. 3. Citrate and pyrophosphate are competitive inhibitors of the enzyme with respect to both 6-phosphogluconate and NADP+. 4. MgCl2 affects the apparent Km for NADP+ at saturating concentrations of 6-phosphogluconate.
MeSH terms
Ammonium Sulfate; Animals; Centrifugation; Chromatography, Affinity; Chromatography, DEAE-Cellulose; Electrophoresis, Polyacrylamide Gel; Kinetics; Magnesium; Mammary Glands, Animal; Molecular Weight; NADP; Phosphogluconate Dehydrogenase; Proteins; Rabbits; Sodium Dodecyl Sulfate
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