Vasoactive intestinal peptide receptor antagonists in rat seminal vesicle membranes.
Vasoactive intestinal peptide (VIP) receptors coupled to activation of adenylate cyclase have been previously identified in seminal vesicle membranes of rat. In the present study we demonstrate that the synthetic peptides [4-Cl-D-Phe6,Leu17]VIP and the growth hormone releasing factor (GRF) analog [Ac-Tyr1,D-Phe2]GRF1-29-NH2 inhibit in a competitive manner the specific 125I-VIP binding to the same membrane preparation. The order of potency of the two peptides compared to VIP was: VIP (IC50 = 1.3 +/- 0.5 nM) greater than [4-Cl-D-Phe6,Leu17]VIP(IC50 = 1600 +/- 45.0 nM) greater than [Ac-Tyr1,D-Phe2]GRF1-29-NH2(IC50 = 290.0 +/- 59.4 nM). Whereas VIP showed a stimulatory activity upon adenylate cyclase with a potency (ED50 = 7.0 +/- 0.7 nM) compatible with the affinity of the VIP binding sites previously described, the other two peptides tested showed no effect at that level. The behavior as antagonists of both [4-Cl-D-Phe6,Leu17]VIP and [Ac-Tyr1,D-Phe2]GRF1-29-NH2 was confirmed by: (a) the parallel shifts of the VIP dose-response curves for stimulation of adenylate cyclase activity in the presence of the antagonists; (b) the close agreement between the binding affinity and the inhibition of adenylate cyclase activity for the two peptides; and (c) the lack of effect of the two antagonists upon the adenylate cyclase activity stimulated by the beta-adrenoceptor agonist isoproterenol which indicates the specificity of the interaction.(ABSTRACT TRUNCATED AT 250 WORDS)
Rodríguez-Pena
M S
MS
Departamento de Bioquímica y Biología Molecular, Universidad de Alcalá, Alcalá de Henares, Madrid, Spain.
Guijarro
L G
LG
Prieto
J C
JC
D000262:Adenylyl Cyclases / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D001667:Binding, Competitive; D013007:Growth Hormone-Releasing Hormone / Q000031:analogs & derivatives / Q000494:pharmacology; D007457:Iodine Radioisotopes; D007545:Isoproterenol / Q000494:pharmacology; D008297:Male; D008566:Membranes / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D011964:Receptors, Gastrointestinal Hormone / Q000037:antagonists & inhibitors* / Q000378:metabolism; D018005:Receptors, Vasoactive Intestinal Peptide; D012669:Seminal Vesicles / Q000187:drug effects* / Q000201:enzymology / Q000648:ultrastructure; D017337:Sermorelin / Q000031:analogs & derivatives; D013268:Stimulation, Chemical; D014660:Vasoactive Intestinal Peptide / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology
D007457:Iodine Radioisotopes; D011964:Receptors, Gastrointestinal Hormone; D018005:Receptors, Vasoactive Intestinal Peptide; C048507:vasoactive intestinal peptide, 4-chloro-Phe(6)-Leu(17)-; D014660:Vasoactive Intestinal Peptide; D017337:Sermorelin; D013007:Growth Hormone-Releasing Hormone; C045616:somatotropin releasing hormone (1-29)-amide, N-Ac-Tyr(1)-Phe(2)-; D000262:Adenylyl Cyclases; D007545:Isoproterenol