Immunocytochemical binding of anti-opsin N-terminal-specific antibodies to the extracellular surface of rod outer segment plasma membranes. Fixation induces antibody binding.
We have examined the binding of anti-opsin antibodies to the plasma membrane of frog retinal rod outer segments (ROS) by fluorescence light microscopy and electron microscopy. Polyclonal and monoclonal antibodies specific for the N-terminal domain of opsin were observed to bind to the extracellular surface of ROS plasma membrane of aldehyde-fixed but not of unfixed retinas. This reaction was found regardless of whether purified ROS, rhodopsin, opsin, or an N-terminal peptide of opsin was used as the immunogen. The fixation-induced binding of these antibodies contrasts with the more frequently noted loss of antigenicity upon fixation. Concanavalin A, however, binds to unfixed ROS plasma membranes. Its binding sites in the plasma membrane may be oligosaccharides in the N-terminal region of opsin. These results suggest that the N-terminal domain of opsin is latent in the native membrane and that changes in conformation may account for its detectability in fixed membranes.
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000378:metabolism*; D000911:Antibodies, Monoclonal; D001360:Avidin; D001710:Biotin; D002462:Cell Membrane / Q000032:analysis; D003208:Concanavalin A / Q000378:metabolism; D005136:Eye Proteins / Q000032:analysis / Q000276:immunology*; D005293:Ferritins; D005404:Fixatives; D005455:Fluorescent Antibody Technique; D005976:Glutaral; D006651:Histocytochemistry / Q000379:methods*; D051379:Mice; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D010446:Peptide Fragments / Q000276:immunology; D010786:Photoreceptor Cells / Q000032:analysis*; D011892:Rana catesbeiana; D012374:Rod Cell Outer Segment / Q000032:analysis*; D017299:Rod Opsins
D000906:Antibodies; D000911:Antibodies, Monoclonal; D005136:Eye Proteins; D005404:Fixatives; D010446:Peptide Fragments; D017299:Rod Opsins; D003208:Concanavalin A; D001360:Avidin; D001710:Biotin; D005293:Ferritins; D005976:Glutaral