title: Water and peptide backbone structure in the active center of bovine rhodopsin.
abstract: Difference FTIR spectra in the conversion of rhodopsin or isorhodopsin to bathorhodopsin were recorded for recombinant wild-type and E113Q bovine rhodopsins. Differences in various vibrational modes between E113Q and the wild-type proteins whose Schiff bases interact with chloride and Glu113, respectively, were analyzed. Water molecules in rhodopsin that change upon formation of bathorhodopsin are detected by a change in frequency of the O-H stretching vibration from 3538 to 3525 cm(-1). This change in the wild-type protein is absent in E113Q. One or a few water molecules are therefore suggested to be located in the proximity of Glu113, the counterion of the Schiff base. Another water vibration at 3564 cm(-1), which is shifted to 3542 cm(-1) in bathorhodopsin in the wild type, persists in E113Q but with approximately 5-cm(-1) shift toward higher frequency. This is due to water molecules that may be located at a site somewhat more remote from Glu113. Structural changes of some peptide carbonyls and amides are also absent in E113Q. On the other hand, the E113Q protein shows shifts of the N-H+ stretching vibrational band, that is probably due to the protonated Schiff base, upon conversion of rhodopsin to bathorhodopsin. No corresponding changes were observed in the wild type. We propose a model in which a water molecule interacts with Glu113, the protonated Schiff base, and peptide carbonyls, and amides. These residues undergo structural changes upon formation of bathorhodopsin.
authors.affiliation: Department of Biophysics, Graduate School of Science, Kyoto University, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Nagata
authors.forename: A
authors.initials: A
authors.lastname: Terakita
authors.forename: H
authors.initials: H
authors.lastname: Kandori
authors.forename: D
authors.initials: D
authors.lastname: Kojima
authors.forename: Y
authors.initials: Y
authors.lastname: Shichida
authors.forename: A
authors.initials: A
authors.lastname: Maeda
mesh_terms: D000818:Animals; D001665:Binding Sites; D002417:Cattle; D018698:Glutamic Acid / Q000235:genetics; D009154:Mutation; D011487:Protein Conformation; D012243:Rhodopsin / Q000031:analogs & derivatives / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012545:Schiff Bases; D017550:Spectroscopy, Fourier Transform Infrared; D014867:Water / Q000737:chemistry
chemical_list: D012545:Schiff Bases; C042882:isorhodopsin; D014867:Water; D018698:Glutamic Acid; C015017:bathorhodopsin; D012243:Rhodopsin