title: Local and distant protein structural changes on photoisomerization of the retinal in bacteriorhodopsin.
abstract: The photoisomerization of the retinal in bacteriorhodopsin is selective and efficient and yields perturbation of the protein structure within femtoseconds. The stored light energy in the primary intermediate is then used for the net translocation of a proton across the membrane in the microsecond to millisecond regime. This study is aimed at identifying how the protein changes on photoisomerization by using the O-H groups of threonines as internal probes. Polarized Fourier-transform IR spectroscopy of [3-(18)O]threonine-labeled and unlabeled bacteriorhodopsin indicates that 3 of the threonines (of a total of 18) change their hydrogen bonding. One is exchangeable in D(2)O, but two are not. A comprehensive mutation study indicates that the residues involved are Thr-89, Thr-17, and Thr-121 (or Thr-90). The perturbation of only three threonine side chains suggests that the structural alteration at this stage of the photocycle is local and specific. Furthermore, the structural change of Thr-17, which is located >11 A from the retinal chromophore, implicates a specific perturbation channel in the protein that accompanies the retinal motion.
authors.affiliation: Department of Biophysics, Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan. kandori@photo2.biophys.kyoto-u.ac.jp
authors.forename: H
authors.initials: H
authors.lastname: Kandori
authors.forename: N
authors.initials: N
authors.lastname: Kinoshita
authors.forename: Y
authors.initials: Y
authors.lastname: Yamazaki
authors.forename: A
authors.initials: A
authors.lastname: Maeda
authors.forename: Y
authors.initials: Y
authors.lastname: Shichida
authors.forename: R
authors.initials: R
authors.lastname: Needleman
authors.forename: J K
authors.initials: JK
authors.lastname: Lanyi
authors.forename: M
authors.initials: M
authors.lastname: Bizounok
authors.forename: J
authors.initials: J
authors.lastname: Herzfeld
authors.forename: J
authors.initials: J
authors.lastname: Raap
authors.forename: J
authors.initials: J
authors.lastname: Lugtenburg
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001436:Bacteriorhodopsins / Q000737:chemistry* / Q000378:metabolism; D019617:Halobacterium salinarum / Q000378:metabolism; D007536:Isomerism; D007700:Kinetics; D008027:Light; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D010777:Photochemistry; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry; D012172:Retinaldehyde / Q000737:chemistry* / Q000378:metabolism; D017550:Spectroscopy, Fourier Transform Infrared; D013912:Threonine
chemical_list: D011994:Recombinant Proteins; D013912:Threonine; D001436:Bacteriorhodopsins; D012172:Retinaldehyde