Subtype-selective immunoprecipitation of the beta 2-adrenergic receptor.
Most antibodies known to interact with beta-adrenergic receptors do not exhibit subtype selectivity, nor do they provide quantitative immunoprecipitation. A monoclonal antibody, G27.1 raised against a synthetic peptide corresponding to the C-terminus of the beta 2-adrenergic receptor of hamster, is selective for the beta 2 subtype. G27.1 provides nearly quantitative immunoprecipitation of the beta 2-adrenergic receptor from hamster lung that has been photoaffinity-labeled and solubilized with sodium dodecyl sulfate. Immunoprecipitation is completely blocked by nanomolar concentrations of the immunizing peptide. This antibody interacts with beta 2-adrenergic receptors from three rodent species, but not with those from humans. When C6 glioma cells, which contain both beta 1- and beta 2-adrenergic receptors, are photoaffinity-labeled in the absence or presence of subtype-selective antagonists, subtype-selective photoaffinity-labeling results. G27.1 can immunoprecipitate beta 2-, but not beta 1-, adrenergic receptors from these cells. Similar results were obtained following subtype-selective photoaffinity-labeling of membranes from rat cerebellum and cerebral cortex. The beta-adrenergic receptors from C6 glioma cells and rat cerebral cortex exist as a mixture of two molecular weight species. These species differ in glycosylation, as shown by endoglycosidase F digestion of crude and immunoprecipitated receptors.
Romano
C
C
Department of Pharmacology, University of Pennsylvania School of Medicine, Philadelphia 19104-6084.
Williams
W V
WV
Fischberg
D J
DJ
Cocero
N
N
Weiner
D B
DB
Greene
M I
MI
Molinoff
P B
PB
D000345:Affinity Labels; D000818:Animals; D000911:Antibodies, Monoclonal; D002531:Cerebellum / Q000032:analysis; D002540:Cerebral Cortex / Q000032:analysis; D003978:Diazomethane / Q000031:analogs & derivatives; D006168:Guinea Pigs; D007114:Immunization; D020120:Iodocyanopindolol / Q000031:analogs & derivatives*; D008168:Lung / Q000032:analysis; D008970:Molecular Weight; D010446:Peptide Fragments / Q000276:immunology; D010869:Pindolol / Q000031:analogs & derivatives; D011233:Precipitin Tests; D051381:Rats; D011943:Receptors, Adrenergic, beta / Q000302:isolation & purification*; D014407:Tumor Cells, Cultured / Q000378:metabolism
D000345:Affinity Labels; D000911:Antibodies, Monoclonal; D010446:Peptide Fragments; D011943:Receptors, Adrenergic, beta; C058075:iodocyanopindolol diazirine; D003978:Diazomethane; D020120:Iodocyanopindolol; D010869:Pindolol