Prostaglandin A1 metabolism and inhibition of cyclic AMP extrusion by avian erythrocytes.
Prostaglandins (PG) inhibit active cyclic AMP export from pigeon red cells, PGA1 and PGA2 most potently (Brunton, L.L., and Mayer, S.E. (1979) J. Biol. Chem. 254, 9714-9720). To probe the mechanism of this action of PGA1, we have studied the interaction of [3H]PGA1 with suspensions of pigeon red cells. The interaction of PGA1 with pigeon red cells is a multistep process of uptake, metabolism, and secretion. [3H] PGA1 rapidly enters red cells and is promptly metabolized (Vmax greater than or equal to 1 nmol/min/10(7) cells) to a compound(s) that remains in the aqueous layer after ethylacetate extraction. The glutathione-depleting agent, diamide, inhibits formation of the PGA1 metabolite. In agreement with the order of potency of other prostaglandins to inhibit cAMP efflux (A much greater than E congruent to B greater than F), PGA2 forms a polar adduct whereas prostaglandins E2, B1, and F2 alpha do not. The red cells secrete the polar metabolite of PGA1 by a saturable mechanism (at 37 degrees C, Km congruent to 0.6 microM, Vmax congruent to 0.5 pmol/min/10(7) cells) that lowered temperatures inhibit (Eact congruent to 21 kcal/mol). Because uptake and metabolism progress with much greater rates than metabolite secretion, red cells transiently concentrate the polar compound intracellularly. Onset and reversal of inhibition of cyclic AMP export by PGA1 coincide with accumulation and secretion of PGA1 metabolite, suggesting that the polar metabolite acts at an intracellular site to inhibit cyclic AMP efflux. In the accompanying Appendix, we present chromatographic and amino acid analyses demonstrating that the polar metabolite is a glutathione adduct of PGA1.
D000818:Animals; D010856:Columbidae; D000242:Cyclic AMP / Q000097:blood*; D004912:Erythrocytes / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism; D066298:In Vitro Techniques; D007700:Kinetics; D008297:Male; D011454:Prostaglandins A / Q000378:metabolism* / Q000494:pharmacology; D013535:Suspensions; D013696:Temperature; D014316:Tritium
D011454:Prostaglandins A; D013535:Suspensions; D014316:Tritium; D000242:Cyclic AMP; D005978:Glutathione; C100573:prostaglandin A1