Improvement of cytochrome P450c17 catalytic processivity as a novel mechanism to attenuate hormonal consequences of enzyme decay in the testes after a gonadotropin surge.
OBJECTIVE
The aim of this study was to gain understanding of the apparent discrepancy between the moderate restriction of testosterone synthesizing capacity and the nearly complete decay of the androgen-producing enzyme, cytochrome P450c17 (CYP17; steroid 17 alpha-hydroxylase/17,20-lyase), in rat testes during the desensitization phase induced by a single, high-dose gonadotropin (human chorionic gonadotropin, hCG) injection.


METHODS
Adult male rats received 25 IU hCG i.v., and purified Leydig cells and crude interstitial cell microsomes were prepared 0, 4, 12, 24, 48, 72, 120 and 192 h afterwards. Component CYP17 activities, i.e. simultaneously catalyzed productive androgen formation and abortive 17 alpha-hydroxyprogesterone release, and their ratio (processivity), were compared with CYP17 levels and testosterone secretion rates.


RESULTS
Leydig cells isolated 48 h after the artificial hCG surge produce 62% less testosterone than control cells upon stimulation in vitro, though CYP17 levels are reduced by 97%. Its total activity decreases by 87%, resulting in a 4.5-fold rise in the turnover number; the processivity is additionally improved 5-fold over controls. Parallel changes occur in interstitial cell microsomes; a negative linear correlation exists between the ratio of productive over total CYP17 activities and the actual CYP17 concentrations. CYP17 is partly denatured to P420 during hCG action, but other heme proteins (cytochrome b5) remain unchanged. Animal treatment with estradiol results in CYP17 down-regulation without any concomitant effect on enzyme processivity.


CONCLUSIONS
Improved CYP17 processivity is suggested to be the consequence of (otherwise rate-limiting) improved electron transfer efficiency towards CYP17. It explains the relatively high testosterone secretion during Leydig cell desensitization and is interpreted to be a protective mechanism to confine adverse consequences of enzyme decay.
Kühn-Velten
W N
WN
Department of Obstetrics and Gynecology, Heinrich Heine University, Düsseldorf, Germany.
D019326:17-alpha-Hydroxyprogesterone / Q000032:analysis / Q000378:metabolism; D000735:Androstenedione / Q000378:metabolism / Q000502:physiology; D000818:Animals; D002478:Cells, Cultured; D006063:Chorionic Gonadotropin / Q000494:pharmacology*; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004721:Endoplasmic Reticulum / Q000201:enzymology; D007985:Leydig Cells / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D008297:Male; D008861:Microsomes / Q000201:enzymology / Q000502:physiology; D011374:Progesterone / Q000378:metabolism / Q000502:physiology; D051381:Rats; D017208:Rats, Wistar; D013254:Steroid 17-alpha-Hydroxylase / Q000032:analysis / Q000378:metabolism*; D013737:Testis / Q000166:cytology / Q000201:enzymology* / Q000378:metabolism*; D013739:Testosterone / Q000032:analysis / Q000378:metabolism; D013997:Time Factors
D006063:Chorionic Gonadotropin; D013739:Testosterone; D000735:Androstenedione; D011374:Progesterone; D019326:17-alpha-Hydroxyprogesterone; D003577:Cytochrome P-450 Enzyme System; D013254:Steroid 17-alpha-Hydroxylase