Substrate specificity of recombinant steroid 21-hydroxylase H. sapiens and the effect of cytochrome b5 on its activity
Abstract
Human cytochrome CYP21 (steroid 21-hydroxylase) has been expressed in Escherichia coli DH5α and the expression level was 150 nmol from 1 liter of bacterial culture. Utilizing two steps of column chromatography has allowed to obtain highly purified protein. Steroids have been screened as potential substrates for the enzyme. CYP21 showed typical spectral change (type I of spectral responce) when binding with progesterone, 17α-hydroxyprogesterone and 11-hydroxyprogesterone. In addition, it has been shown that cytochrome b5 does not affect hydroxylase activity of CYP21 toward progesterone, 17a-hydroxyprogesterone and 11-hydroxyprogesterone.
About the Authors
V. A. Kopats
Институт биоорганической химии НАН Беларуси
Belarus
Belarus
G. V. Sergeev
Институт биоорганической химии НАН Беларуси
Belarus
Belarus
A. A. Gilep
Институт биоорганической химии НАН Беларуси
Belarus
Belarus
S. A. Usanov
Институт биоорганической химии НАН Беларуси
Belarus
Belarus
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