title: Photoactivation of rhodopsin involves alterations in cysteine side chains: detection of an S-H band in the Meta I-->Meta II FTIR difference spectrum.
abstract: FTIR difference spectroscopy has been used to study the role of cysteine residues in the photoactivation of rhodopsin. A positive band near 2550 cm-1 with a low frequency shoulder is detected during rhodopsin photobleaching, which is assigned on the basis of its frequency and isotope shift to the S-H stretching mode of one or more cysteine residues. Time-resolved studies at low temperature show that the intensity of this band correlates with the formation and decay kinetics of the Meta II intermediate. Modification of rhodopsin with the reagent NEM, which selectively reacts with the SH groups of Cys-140 and Cys-316 on the cytoplasmic surface of rhodopsin, has no effect on the appearance of this band. Four other cysteine residues are also unlikely to contribute to this band because they are either thio-palmitylated (Cys-322 and Cys-323) or form a disulfide bond (Cys-110 and Cys-187). On this basis, it is likely that at least one of the four remaining cysteine residues in rhodopsin is structurally active during rhodopsin photoactivation. The possibility is also considered that this band arises from a transient cleavage of the disulfide bond between cysteine residues 110 and 187.
authors.affiliation: Department of Physics, Boston University, Massachusetts 02215.
authors.forename: P
authors.initials: P
authors.lastname: Rath
authors.forename: P H
authors.initials: PH
authors.lastname: Bovee-Geurts
authors.forename: W J
authors.initials: WJ
authors.lastname: DeGrip
authors.forename: K J
authors.initials: KJ
authors.lastname: Rothschild
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D003545:Cysteine; D004220:Disulfides; D005033:Ethylmaleimide; D012243:Rhodopsin / Q000737:chemistry* / Q000528:radiation effects; D012374:Rod Cell Outer Segment / Q000378:metabolism; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods
chemical_list: D004220:Disulfides; D012243:Rhodopsin; D003545:Cysteine; D005033:Ethylmaleimide