Purification of 20 alpha-hydroxysteroid oxidoreductase from bovine fetal erythrocytes.
NADPH-dependent 20 alpha-hydroxysteroid oxidoreductase (20 alpha-HSD; EC 1.1.1.149) from bovine fetal erythrocytes was obtained for the first time free of hemoglobin by a new 2,500-fold purification scheme. This was achieved by a sequence of calcium phosphate gel absorption, ammonium sulfate fractionation, and affinity chromatography. The present results lead us to believe that the NADPH-dependent 3 beta-hydroxysteroid oxidoreductase activity, which was co-purified with 20 alpha-activity, may originate at the active site of 20 alpha-HSD (2).
D015089:20-Hydroxysteroid Dehydrogenases / Q000302:isolation & purification*; D000645:Ammonium Sulfate; D000818:Animals; D002130:Calcium Phosphates; D002417:Cattle; D002846:Chromatography, Affinity; D004912:Erythrocytes / Q000201:enzymology*; D005260:Female; D005312:Fetal Blood / Q000201:enzymology*; D011247:Pregnancy
D002130:Calcium Phosphates; C485828:alpha-tricalcium phosphate; C485830:tetracalcium phosphate; C485838:calcium phosphate, monobasic, anhydrous; C020243:calcium phosphate; D015089:20-Hydroxysteroid Dehydrogenases; C485829:calcium phosphate, dibasic, anhydrous; D000645:Ammonium Sulfate