Succinate stimulation of isocitrate supported deoxycorticosterone metabolism in rat adrenal mitochondria by a synergistic mechanism.
The relative effectiveness of succinate and isocitrate in supplying NADPH for cholesterol and deoxycorticosterone (DOC) metabolism by rat adrenal mitochondria has been investigated. As previously seen for cholesterol metabolism, isocitrate supported DOC metabolism at a higher rate than succinate. Maximal rates of DOC metabolism, however, required 10 times more precursor (10mM) than cholesterol metabolism. Addition of DOC to mitochondria inhibited cholesterol metabolism, indicating competition for NADPH between these pathways. Coaddition of these reducing precursors resulted in a substantially greater than additive rate of DOC hydroxylation, but not cholesterol metabolism. The synergistic effect was seen with both 11 beta- and 18 hydroxylation. For both precursors, the synergism was maximal upon addition of only 1 mM of the second precursor. The synergistic effect was far more resistant to added KCN and malonate than succinate supported DOC metabolism, and neither inhibitor affected isocitrate supported DOC metabolism. These results suggest that while cytochromes P450scc and P450(11 beta) use a common supply of NADPH generated by each precursor, there is a pool of NADPH that is only effectively synthesised upon coaddition of precursors and only utilised by cytochrome P450(11 beta). This second NADPH pool may be produced in response to potentiated isocitrate dehydrogenase activity or activation of a different NADPH generating system.
McNamara
B C
BC
Dept. of Pharmacology, University of Wisconsin Medical School, Madison 53705.
Jefcoate
C R
CR
D000311:Adrenal Glands / Q000378:metabolism* / Q000648:ultrastructure; D000818:Animals; D002784:Cholesterol / Q000502:physiology; D003900:Desoxycorticosterone / Q000378:metabolism*; D004357:Drug Synergism; D007523:Isocitrates / Q000494:pharmacology*; D008928:Mitochondria / Q000378:metabolism* / Q000648:ultrastructure; D009249:NADP / Q000378:metabolism; D011863:Radioimmunoassay; D051381:Rats; D013386:Succinates / Q000494:pharmacology*
D007523:Isocitrates; D013386:Succinates; D003900:Desoxycorticosterone; D009249:NADP; D002784:Cholesterol