title: Direct observation of the structural change of Tyr174 in the primary reaction of sensory rhodopsin II.
abstract: Sensory rhodopsin II (SRII) is a negative phototaxis receptor containing retinal as its chromophore, which mediates the avoidance of blue light. The signal transduction is initiated by the photoisomerization of the retinal chromophore, resulting in conformational changes of the protein which are transmitted to a transducer protein. To gain insight into the SRII sensing mechanism, we employed time-resolved ultraviolet resonance Raman spectroscopy monitoring changes in the protein structure in the picosecond time range following photoisomerization. We used a 450 nm pump pulse to initiate the SRII photocycle and two kinds of probe pulses with wavelengths of 225 and 238 nm to detect spectral changes in the tryptophan and tyrosine bands, respectively. The observed spectral changes of the Raman bands are most likely due to tryptophan and tyrosine residues located in the vicinity of the retinal chromophore, i.e., Trp76, Trp171, Tyr51, or Tyr174. The 225 nm UVRR spectra exhibited bleaching of the intensity for all the tryptophan bands within the instrumental response time, followed by a partial recovery with a time constant of 30 ps and no further changes up to 1 ns. In the 238 nm UVRR spectra, a fast recovering component was observed in addition to the 30 ps time constant component. A comparison between the spectra of the WT and Y174F mutant of SRII indicates that Tyr174 changes its structure and/or environment upon chromophore photoisomerization. These data represent the first real-time observation of the structural change of Tyr174, of which functional importance was pointed out previously.
authors.affiliation: Department of Chemistry, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka 560-0043, Japan.
authors.forename: Misao
authors.initials: M
authors.lastname: Mizuno
authors.forename: Yuki
authors.initials: Y
authors.lastname: Sudo
authors.forename: Michio
authors.initials: M
authors.lastname: Homma
authors.forename: Yasuhisa
authors.initials: Y
authors.lastname: Mizutani
mesh_terms: D006216:Halobacteriaceae; D025602:Halorhodopsins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D025603:Sensory Rhodopsins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D013059:Spectrum Analysis, Raman; D013997:Time Factors; D014443:Tyrosine / Q000737:chemistry*; D014732:Vibration
chemical_list: D025602:Halorhodopsins; D025603:Sensory Rhodopsins; C061508:sensory rhodopsin II protein, archaeal; D014443:Tyrosine