Premium
Structure and stereochemistry of products of hydroxylation of human steroid hormones by a housefly cytochrome P450 (CYP6A1)
Author(s) -
Jacobsen Neil E.,
Kövér Katalin E.,
Murataliev Marat B.,
Feyereisen René,
Walker F. Ann
Publication year - 2006
Publication title -
magnetic resonance in chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.483
H-Index - 72
eISSN - 1097-458X
pISSN - 0749-1581
DOI - 10.1002/mrc.1767
Subject(s) - chemistry , heteronuclear molecule , heteronuclear single quantum coherence spectroscopy , stereochemistry , two dimensional nuclear magnetic resonance spectroscopy , hydroxylation , cytochrome p450 , steroid , cytochrome b5 , nuclear magnetic resonance spectroscopy , enzyme , biochemistry , hormone
The structure and stereochemistry of nine steroid metabolites isolated in quantities ranging from 0.15 to 1.8 mg were determined using a variety of NMR techniques, including heteronuclear multiple bond correlation (HMBC) using broadband adiabatic 13 C pulses and phase‐sensitive data presentation. Testosterone, androstenedione and progesterone were oxidized with housefly cytochrome P450 6A1 enzyme reconstituted in vitro with housefly NADPH cytochrome P450 reductase and cytochrome b 5 . NMR analysis in CD 3 OD using a modified HMBC sequence as well as 2D heteronuclear single quantum correlation (HSQC), COSY and nuclear Overhauser and exchange spectroscopy (NOESY), combined with a detailed analysis of J couplings showed that hydroxylation occurs exclusively on the β‐face of the steroids, at positions 2, 12, and 15. Copyright © 2006 John Wiley & Sons, Ltd.