Indirect immunofluorescence localization of beta-adrenergic receptors and G-proteins in human A431 cells.
Polyclonal antibodies directed against (i) rodent lung beta 2-adrenergic receptor, (ii) a synthetic fragment of an extracellular domain of the receptor, and (iii) human placenta G-protein beta-subunits, were used to localize these antigens in situ in intact and permeabilized human epidermoid carcinoma A431 cells. Antibodies directed against beta 2-adrenergic receptors showed a punctate immunofluorescence staining throughout the cell surface of fixed intact cells. Punctate staining was also observed in clones of Chinese hamster ovary cells transfected with an expression vector harbouring the gene for the hamster beta 2-adrenergic receptor. The immunofluorescence observed with anti-receptor antibodies paralleled the level of receptor expression. In contrast, the beta-subunits common to G-proteins were not stained in fixed intact cells, presumably reflecting their intracellular localization. In detergent-permeabilized fixed cells, strong punctate staining of G beta-subunits was observed throughout the cytoplasm. This is the first indirect immunofluorescence localization of beta-adrenergic receptors and G-proteins. Punctate immunofluorescence staining suggests that both antigens are distributed in clusters.
Wang
H Y
HY
Department of Pharmacology, State University of New York, Stony Brook 11794-8651.
Berrios
M
M
Malbon
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CC
D000818:Animals; D000906:Antibodies; D000941:Antigens / Q000276:immunology; D002294:Carcinoma, Squamous Cell; D002462:Cell Membrane / Q000032:analysis; D006224:Cricetinae; D003593:Cytoplasm / Q000032:analysis; D005455:Fluorescent Antibody Technique; D019204:GTP-Binding Proteins / Q000032:analysis* / Q000276:immunology; D006801:Humans; D015151:Immunoblotting; D020120:Iodocyanopindolol; D051379:Mice; D010869:Pindolol / Q000031:analogs & derivatives / Q000378:metabolism; D010920:Placenta / Q000032:analysis; D011943:Receptors, Adrenergic, beta / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000906:Antibodies; D000941:Antigens; D011943:Receptors, Adrenergic, beta; D020120:Iodocyanopindolol; D010869:Pindolol; D019204:GTP-Binding Proteins