Abstract
The apoenzyme and holoenzyme (NADP
+ complex) of human placental 17β-hydroxysteroid dehydrogenase (17β-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 17β-HSD—NADP
+ complex was prepared using NADP
+ elution in a Blue-Sepharose column. The two forms have different
A
280/
A
260 ratios and are suitable for further study of enzyme—cofactor interactions.