Characterization of vasoactive intestinal peptide receptors in rat seminal vesicle.
Receptors for vasoactive intestinal peptide (VIP) in membranes from rat seminal vesicle were examined using 125I-labeled VIP as ligand. The receptor binding was rapid, reversible, saturable, specific, and dependent on temperature and membrane concentration. At 15 degrees C, the stoichiometric data suggested the presence of two classes of VIP receptors with Kd values of 0.54 and 44.4 nM and binding capacities of 73 and 1,065 fmol VIP/mg membrane protein, respectively. The interaction showed a high degree of specificity, as suggested by competition experiments with various peptides structurally related to VIP as follows: helodermin was 10 times, secretin 30 times, and rat growth hormone-releasing factor 300 times less potent than VIP, whereas glucagon did not recognize VIP receptors in concentrations of up to 10 microM. The binding of 125I-VIP to membranes was sensitive to the presence of GTP in the incubation medium in a dose-dependent manner. To characterize the molecular weight of these VIP receptors, 125I-VIP was covalently bound to membranes from rat seminal vesicle using dithiobis(succinimidyl propionate); sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the solubilized receptor revealed the presence of a specific component with a molecular mass of 47,000 Da as estimated in denaturing conditions. These findings, together with the known presence of VIP-containing nerves in the seminal vesicle, suggest a direct physiological role for this peptide in this accessory gland of the male genital tract.
Guijarro
L G
LG
Departamento de Bioquímica y Biología Molecular, Universidad de Alcalá, Alcalá de Henares-Madrid, Spain.
Rodriguez-Pena
M S
MS
Prieto
J C
JC
D000818:Animals; D001667:Binding, Competitive; D002462:Cell Membrane / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006150:Guanine Nucleotides / Q000494:pharmacology; D007700:Kinetics; D008297:Male; D008970:Molecular Weight; D051381:Rats; D011919:Rats, Inbred Strains; D011964:Receptors, Gastrointestinal Hormone / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D018005:Receptors, Vasoactive Intestinal Peptide; D012669:Seminal Vesicles / Q000378:metabolism*; D014660:Vasoactive Intestinal Peptide / Q000378:metabolism*
D006150:Guanine Nucleotides; D011964:Receptors, Gastrointestinal Hormone; D018005:Receptors, Vasoactive Intestinal Peptide; D014660:Vasoactive Intestinal Peptide