Expression of VIP receptors in mouse peritoneal macrophages: functional and molecular characterization.
Receptors for VIP in mouse peritoneal macrophages (MPM) were examined using [125I]labeled VIP as ligand. The receptor binding was rapid, reversible, saturable, specific, and dependent on time, pH, temperature and cell concentration. At 15 degrees C, the stoichiometric data suggested the presence of two classes of VIP receptors with Kd values of 1.05 +/- 0.2 and 66.4 +/- 11.0 nM and binding capacities of 19.2 +/- 2.8 and 706.6 +/- 172.0 fmol VIP/10(6) cells. The interaction showed a high degree of specificity, as suggested by competition experiments with various peptides structurally related to VIP as follows: VIP > helodermin > rGRF > PHI >> secretin. Glucagon, pancreastatin, somatostatin, insulin, and octapeptide of cholecystokinin (CCK 26-33) were ineffective at concentrations as high as 1 microM. VIP was a potent and efficient stimulator of cyclic AMP production in MPM. The stimulation was observed at a concentration as low as 0.01 nM VIP. Half-maximal stimulation (ED50) was observed at 1.0 +/- 0.2 nM VIP, and maximal stimulation (three-fold above basal levels) was obtained between 0.1-1 microM. The cyclic AMP system of mouse peritoneal macrophages showed a high specificity for VIP. The order of potency observed in inducing cyclic AMP production was VIP > helodermin > rGRF > PHI >> secretin. Glucagon, insulin, pancreastatin, somatostatin and octapeptide of cholecystokinin did not modify cyclic AMP levels at concentrations as high as 1 microM.(ABSTRACT TRUNCATED AT 250 WORDS)
Calvo
J R
JR
Departamento de Bioquímica Médica y Biología Molecular, Facultad de Medicina, Universidad de Sevilla, Spain.
Montilla
M L
ML
Guerrero
J M
JM
Segura
J J
JJ
D000818:Animals; D000242:Cyclic AMP / Q000096:biosynthesis; D017737:Macrophages, Peritoneal / Q000737:chemistry*; D051379:Mice; D008970:Molecular Weight; D018005:Receptors, Vasoactive Intestinal Peptide / Q000032:analysis* / Q000502:physiology; D012680:Sensitivity and Specificity; D014660:Vasoactive Intestinal Peptide / Q000378:metabolism / Q000494:pharmacology
D018005:Receptors, Vasoactive Intestinal Peptide; D014660:Vasoactive Intestinal Peptide; D000242:Cyclic AMP