Inhibition of (+)[3H]SKF 10,047 binding to rat brain membranes by FAB fragments from a monoclonal antibody directed against the opioid receptor.
Examination of the binding of (+)[3H]SKF 10,047 to rat brain membranes indicated that at a low concentration most of the binding was to the haloperidol-sensitive binding site. Titration curves exhibited a displacement potency order of haloperidol greater than (+)SKF 10,047 = 1,3-diorthotolyl-guanidine much much greater than (-)SKF 10,047 much greater than phencyclidine analogues. The effect of Fab fragments from a monoclonal antibody, OR-689.2.4, directed against the opioid receptor on the binding of (+)[3H]SKF 10,047 to rat brain membranes was examined. The specificity of this antibody for the opioid receptor has been determined by its ability to inhibit the binding of mu and delta opioid peptides to rat brain but not the binding of kappa opioid ligands or nonopioid ligands specific for other receptors. The Fab fragments blocked and displaced specifically bound (+)[3H]SKF 10,047 in a titratable manner. Increasing the incubation time of the membranes with the Fab fragments increased the percent inhibition obtained. The Fab fragments acted as noncompetitive inhibitors of (+)[3H]SKF 10,047 binding. A (+)SKF 10,047 binding site in rat brain appears to share a common structural domain with mu and delta opioid receptors.
Tyler
C B
CB
Department of Pharmacology, University of Rochester, School of Medicine and Dentistry, New York 14642.
Bidlack
J M
JM
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism*; D001665:Binding Sites; D001921:Brain / Q000378:metabolism*; D007128:Immunoglobulin Fragments / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D010620:Phenazocine / Q000031:analogs & derivatives* / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D011957:Receptors, Opioid / Q000276:immunology* / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism
D000911:Antibodies, Monoclonal; D007128:Immunoglobulin Fragments; D011957:Receptors, Opioid; C012134:SK&F 10047; D010620:Phenazocine