title: Identification of FTIR bands due to internal water molecules around the quinone binding sites in the reaction center from Rhodobacter sphaeroides.
abstract: The bacterial reaction center (RC) is a membrane protein complex that performs photosynthetic electron transfer from a bacteriochlorophyll dimer to quinone acceptors Q(A) and Q(B). Q(B) accepts electrons from the primary quinone, Q(A), in two sequential electron transfer reactions coupled to uptake of a proton from solution. It has been suggested that water molecules along the proton uptake pathway are protonated upon quinone reduction on the basis of FTIR difference spectra [Breton, J., and Nabedryk, E. (1998) Photosynth. Res. 55, 301-307]. We examined the possible involvement of water molecules in the photoreaction processes by studying (18)O water isotope effects on FTIR difference spectra resulting from formation of Q(A)(-) and Q(B)(-). Continuum bands in D(2)O due to Q(B)(-) formation in the 2300-1800 cm(-1) region did not show spectral shifts by (18)O water in the wild-type (WT) RC, suggesting that these bands do not originate from (protonated) water. In contrast, the Q(B)(-)/Q(B) spectrum of the EQ-L212 mutant RC showed a spectral shift of a band near 2100 cm(-1) due to (18)O water substitution, consistent with protonation of internal water. FTIR shifts due to (18)O water were also observed following formation of Q(A)(-) and Q(B)(-) in the spectral region of 3700-3500 cm(-1) characteristic of weakly hydrogen bonded water. The water responsible for the Q(B)(-) change was localized near Glu-L212 by spectral shifts in mutant RCs. The weakly hydrogen bonded water perturbed by quinone reduction may play a role in stabilizing the charge-separated state.
authors.affiliation: Department of Frontier Materials, Nagoya Institute of Technology, Showa-ku, Nagoya 466-8555, Japan.
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Iwata
authors.forename: Mark L
authors.initials: ML
authors.lastname: Paddock
authors.forename: Melvin Y
authors.initials: MY
authors.lastname: Okamura
authors.forename: Hideki
authors.initials: H
authors.lastname: Kandori
mesh_terms: D019943:Amino Acid Substitution / Q000528:radiation effects; D001665:Binding Sites; D008027:Light; D009154:Mutation / Q000235:genetics; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry*; D011809:Quinones / Q000737:chemistry*; D012242:Rhodobacter sphaeroides / Q000737:chemistry* / Q000528:radiation effects; D017550:Spectroscopy, Fourier Transform Infrared; D014732:Vibration; D014867:Water / Q000737:chemistry*
chemical_list: D045322:Photosynthetic Reaction Center Complex Proteins; D011809:Quinones; D014867:Water