Bovine adrenal cytochrome P-450(11 beta)-mediated conversion of 11-deoxycortisol to 18- and 19-hydroxy derivatives; structural analysis by 1H-NMR.
Incubation of 11-deoxycortisol with a cytochrome P-450(11 beta)-reconstituted system yielded, in addition to cortisol, several new steroid products. In this study, structures of the three steroid products were elucidated. Retention time of the first product (Peak 2 substance) coincided with that of authentic 18-hydroxycortisol on reverse phase HPLC. To further confirm the chemical identity of this product, the purified sample was subjected to 1H-NMR analysis. The spectrum was essentially identical to that of 18-hydroxycortisol. The retention time of the second product (Peak 3 substance) did not coincide with those of commonly occurring steroids. The one- and two-dimension 1H-NMR spectra provided strong evidence for its structure of 19-hydroxy-11-deoxycortisol. The retention time of the third product (Peak 4 substance) did not coincide with those of commonly occurring steroids. The 1H-NMR spectrum showed the presence of signals of 19-CH3 and 18-CH2 protons. There was also evidence that this product is not hydroxylated at the 11-position. Further analysis of the COSY spectra identified its structure as 18-hydroxy-11-deoxycortisol. From these results, we conclude that bovine P-450(11 beta) can catalyze the hydroxylation of 11-deoxycortisol at 11 beta-, 18- and 19-positions and produce cortisol, 18-hydroxy-11-deoxycortisol, 18-hydroxycortisol and 19-hydroxy-11-deoxycortisol.
Ohta
M
M
Department of Biochemistry, Osaka University Medical School, Japan.
Fujii
S
S
Miura
R
R
Nonaka
Y
Y
Okamoto
M
M
D000302:Adrenal Cortex / Q000737:chemistry / Q000201:enzymology*; D000818:Animals; D002384:Catalysis; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D003350:Cortodoxone / Q000032:analysis / Q000378:metabolism*; D003577:Cytochrome P-450 Enzyme System / Q000502:physiology*; D006854:Hydrocortisone / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism; D006900:Hydroxylation; D009682:Magnetic Resonance Spectroscopy
D003577:Cytochrome P-450 Enzyme System; C033689:18-hydroxycortisol; D003350:Cortodoxone; D006854:Hydrocortisone