Phospholipases modulate the rat testicular androgen biosynthetic pathway in vitro.
The role of membrane phospholipids in testicular androgen biosynthesis was investigated by monitoring the effects of phospholipase treatments on the activities of the steroid transforming enzymes. Androgen biosynthesis in untreated rat testicular microsomes was examined by monitoring the temporal appearance of pregnenolone metabolites and was found to proceed through the 4-ene route. When phospholipase A2 was included, the 5-ene steroids 17-hydroxypregnenolone and dehydroepiandrosterone (DHEA) were formed in greater quantities, and the production of 4-ene steroids was reduced indicating that the conversion of 5-ene steroids to the 4-ene configuration was inhibited by phospholipase A2 treatment. Phospholipase C, in addition to inhibiting this step, also inhibited the conversion of C21 steroids to C19 steroids. When the enzymatic steps were measured individually, phospholipase A2 inhibited 3 beta-hydroxysteroid dehydrogenase-isomerase (3 beta-HSD-Isomerase) with an ED50 of 73 mU/ml but had no effect on the activities of 17-hydroxylase, C-17, 20 lyase, or 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD). However, though phospholipase C treatment inhibited 3 beta-HSD-Isomerase, it caused less inhibition (the ED50 value was 149 mU/ml). Furthermore, 17-hydroxylase and C-17, 20 lyase activities were also inhibited by phospholipase C treatment (ED50 values were 410 and 343 mU/ml, respectively), but no effect on 17 beta-HSD was observed. The differences in the apparent phospholipid requirements of the steroidogenic enzymes provides the possibility that the metabolic fate of pregnenolone may be regulated by changes in the phospholipid composition of the microenvironment.
Cooke
G M
GM
Department of Pharmacology, McGill University, Montreal, Quebec, Canada.
Robaire
B
B
D015067:17-Hydroxysteroid Dehydrogenases / Q000378:metabolism; D000728:Androgens / Q000096:biosynthesis*; D000818:Animals; D006863:Hydrogen-Ion Concentration; D008297:Male; D008861:Microsomes / Q000378:metabolism; D009097:Multienzyme Complexes / Q000378:metabolism; D009243:NAD / Q000032:analysis; D009249:NADP / Q000032:analysis; D010740:Phospholipases / Q000502:physiology*; D010741:Phospholipases A / Q000502:physiology*; D054467:Phospholipases A2; D011284:Pregnenolone / Q000378:metabolism; D011375:Progesterone Reductase / Q000378:metabolism; D011506:Proteins / Q000032:analysis; D051381:Rats; D011919:Rats, Inbred Strains; D013254:Steroid 17-alpha-Hydroxylase / Q000378:metabolism; D013251:Steroid Isomerases / Q000378:metabolism; D013737:Testis / Q000378:metabolism*; D010738:Type C Phospholipases / Q000502:physiology*
C021425:3 beta-hydroxysteroid oxidoreductase-delta(5) 3-ketosteroid isomerase; D000728:Androgens; D009097:Multienzyme Complexes; D011506:Proteins; D009243:NAD; D009249:NADP; D011284:Pregnenolone; D015067:17-Hydroxysteroid Dehydrogenases; D011375:Progesterone Reductase; C032690:3-alpha-(17-beta)-hydroxysteroid dehydrogenase (NAD(+)); D013254:Steroid 17-alpha-Hydroxylase; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2; D010738:Type C Phospholipases; D013251:Steroid Isomerases