Biochemical aspects of the visual process. XXVIII. Classification of sulfhydryl groups in phodopsin and other photoreceptor membrane proteins.
Reaction of isolated bovine rod outer segment membrane with radioactive N-ethylmaleimide, both in the presence and absence of 1% dodecyl sulfate followed by dodecyl sulfate-polyacrylamide gel electrophoresis, shows that six sulfhydryl groups (96% of total sulfhydryl in this membrane) are located on the rhodopsin molecule. On the basis of their reactivity towards rho-chloromercuribenzoate and rho-chloromercuribenzene sulfonate in suspensions of outer segment membranes, the sulfhydryl groups of rhodopsin can be divided into three pairs. One pair is rapidly modified, both in light and darkness. This modification does not impair the recombination capacity of opsin with 11-cis retinaldehyde under regeneration of rhodopsin. A second pair is modified upon prolonged interaction with the rho-chloromercuriderivatives in darkness. Modification of this pair leaves the typical rhodopsin absorbance at 500 nm intact, but a proportional loss of recombination capacity does occur. The third pair is only modified after illumination and isprobably located in the vicinity of the chromophoric center. The differences between these results and those obtained by modification with dithiobis-(2-nitrobenzoic acid) or N-ethylmaleimide in suspension, where even upon prolonged exposure to light as well as in darkness only two sulfhydryl groups of rhodopsin are modified, is explained by the detergent-like character of the rho-chloromercuri-derivatives.
D000818:Animals; D001665:Binding Sites; D002417:Cattle; D002462:Cell Membrane / Q000032:analysis; D002729:Chloromercuribenzoates; D004228:Dithionitrobenzoic Acid; D004591:Electrophoresis, Polyacrylamide Gel; D005033:Ethylmaleimide; D007700:Kinetics; D009419:Nerve Tissue Proteins; D010786:Photoreceptor Cells / Q000032:analysis*; D011485:Protein Binding; D012168:Retinal Pigments; D012243:Rhodopsin; D013438:Sulfhydryl Compounds / Q000032:analysis; D013997:Time Factors; D014461:Ultracentrifugation; D014785:Vision, Ocular
D002729:Chloromercuribenzoates; D009419:Nerve Tissue Proteins; D012168:Retinal Pigments; D013438:Sulfhydryl Compounds; D012243:Rhodopsin; D004228:Dithionitrobenzoic Acid; D005033:Ethylmaleimide