Preparation of the apoenzyme and holoenzyme forms of human 17 beta-hydroxysteroid dehydrogenase.
The apoenzyme and holoenzyme (NADP+ complex) of human placental 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD) were prepared from affinity chromatography using various elutions by column liquid chromatography. The apoenzyme was obtained using NAD+ elution in a Blue-Sepharose column, followed by NAD+ separation on a Phenyl-Superose hydrophobic-interaction or a Mono Q anion-exchange column. The 17 beta-HSD-NADP+ complex was prepared using NADP+ elution in a Blue-Sepharose column. The two forms have different A280/A260 ratios and are suitable for further study of enzyme-cofactor interactions.
Jin
J Z
JZ
MRC Group in Molecular Endocrinology, CHUL Research Center, Ste-Foy, Quebec, Canada.
Azzi
A
A
Wang
J Y
JY
Lin
S X
SX
D015067:17-Hydroxysteroid Dehydrogenases / Q000302:isolation & purification* / Q000378:metabolism; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007527:Isoenzymes / Q000302:isolation & purification* / Q000378:metabolism; D009243:NAD / Q000302:isolation & purification; D010920:Placenta / Q000201:enzymology; D012685:Sepharose / Q000031:analogs & derivatives
D007527:Isoenzymes; D009243:NAD; C029483:Blue-Sepharose; D012685:Sepharose; D015067:17-Hydroxysteroid Dehydrogenases