The accessibility of bovine rhodopsin in photoreceptor membranes.
Bovine photoreceptor membranes have been treated with proteases to determine the accessibility of rhodopsin to these large, water soluble molecules. The polypeptides that remain associated with the membranous structure after proteolysis were detected by sodium dodecyl sulfate gel electrophoresis. Thermolysin and chymotrypsin degraded rhodopsin (apparent mol wt 35,000-36,000) to fragments of 29,000 and 23,000 apparent mol wt, respectively, without affecting the chromophoric absorption of the molecule or removing the region of the polypeptide carrying carbohydrate. The two fragments were isolated and their amino acid compositions were determined. They do not appear to be more hydrophobic than rhodopsin. Subtilisin, at low concentration and temperature, produced a fragment with the same molecular weight as that produced by thermolysin. At higher concentrations, subtilisin yields major fragments of mol wt 23,000 and 20,000 without affecting the chromophoric absorption. Two intermediate fragments of apparent mol wt 29,000 and 26,000 were detected during the course of this degradation. Carbohydrate is retained by all but the smallest fragment. Bleaching of the photoreceptor pigment did not appreciably alter any of the fragmentation patterns. Trypsin did not alter the molecular weight of rhodopsin under the conditions of this study. Approximately 35-45% of rhodopsin appears to be accessible to the aqueous environment and can be removed without affecting the chromophoric properties of the retinaldehyde-carrying region which remains bound to the membrane.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000032:analysis / Q000648:ultrastructure; D002499:Centrifugation, Density Gradient; D002918:Chymotrypsin; D004591:Electrophoresis, Polyacrylamide Gel; D005613:Freeze Etching; D008854:Microscopy, Electron; D008970:Molecular Weight; D010446:Peptide Fragments; D010761:Phosphorus Radioisotopes; D010786:Photoreceptor Cells / Q000032:analysis* / Q000648:ultrastructure; D012160:Retina / Q000032:analysis; D012168:Retinal Pigments / Q000032:analysis*; D012243:Rhodopsin / Q000032:analysis*; D012967:Sodium Dodecyl Sulfate; D013381:Subtilisins; D013820:Thermolysin; D013997:Time Factors
D000596:Amino Acids; D010446:Peptide Fragments; D010761:Phosphorus Radioisotopes; D012168:Retinal Pigments; D012967:Sodium Dodecyl Sulfate; D012243:Rhodopsin; D013381:Subtilisins; D002918:Chymotrypsin; D013820:Thermolysin