title: Electron-nuclear and electron-electron double resonance spectroscopies show that the primary quinone acceptor QA in reaction centers from photosynthetic bacteria Rhodobacter sphaeroides remains in the same orientation upon light-induced reduction.
abstract: Reaction centers (RCs) from the photosynthetic bacterium Rhodobacter (Rb.) sphaeroides R-26 exhibit changes in the recombination kinetics of the charge-separated radical-pair state, P(·+) Q(A)(·-), composed of the dimeric bacteriochlorophyll donor P and the ubiquinone-10 acceptor Q(A), depending on whether the RCs are cooled to cryogenic temperatures in the dark or under continuous illumination (Kleinfeld et al. Biochemistry 1984, 23, 5780-5786). Structural changes near redox-active cofactors have been postulated to be responsible for these changes in kinetics and to occur in the course of light-induced oxidation and reduction of the cofactors thereby assuring a high quantum yield. Here we investigated such potential light-induced structural changes, associated with the formation of P(·+) Q(A)(·-), via pulsed electron-nuclear double resonance (ENDOR) at Q-band (34 GHz) and pulsed electron-electron double resonance (PELDOR) at W-band (95 GHz). Two types of light excitation have been employed for which identical RC samples were prepared: (a) one sample was frozen in the dark and then illuminated to generate transient P(·+) Q(A)(·-), and (b) one was frozen under illumination which resulted in both trapped and transient P(·+) Q(A)(·-) at 80 K. The hyperfine interactions between Q(A)(·-) and the protein were found to be the same in RCs frozen in the dark as in RCs frozen under illumination. Furthermore, these interactions are completely consistent with those observed in RC crystals frozen in the dark. Thus, QA remains in its binding site with the same position and orientation upon reduction. This conclusion is consistent with the result of our orientation-resolving PELDOR experiments on transient P(·+) Q(A)(·-) radical pairs. However, these findings are incompatible with the recently proposed ~60° reorientation of Q(A) upon its photoreduction, as deduced from an analysis of Q-band quantum-beat oscillations (Heinen et al. J. Am. Chem. Soc. 2007, 129, 15935-15946). Such a large reorientation appears improbable, and our objections against this proposition are substantiated here in detail. Our results show that Q(A) is initially in an orientation that is favorable for its light-driven reduction. This diminishes the reorganization requirements for fast electron reduction and high quantum efficiency.
authors.affiliation: Max-Planck-Institut für Bioanorganische Chemie, 45470 Mülheim an der Ruhr, Germany. mafloresr@asu.edu
authors.forename: Marco
authors.initials: M
authors.lastname: Flores
authors.forename: Anton
authors.initials: A
authors.lastname: Savitsky
authors.forename: Mark L
authors.initials: ML
authors.lastname: Paddock
authors.forename: Edward C
authors.initials: EC
authors.lastname: Abresch
authors.forename: Alexander A
authors.initials: AA
authors.lastname: Dubinskii
authors.forename: Melvin Y
authors.initials: MY
authors.lastname: Okamura
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Lubitz
authors.forename: Klaus
authors.initials: K
authors.lastname: Möbius
mesh_terms: D004578:Electron Spin Resonance Spectroscopy; D004579:Electron Transport; D004583:Electrons; D005609:Free Radicals / Q000737:chemistry; D007700:Kinetics; D008027:Light; D010084:Oxidation-Reduction; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry*; D011789:Quantum Theory; D011809:Quinones / Q000737:chemistry*; D012242:Rhodobacter sphaeroides / Q000737:chemistry* / Q000378:metabolism; D013696:Temperature
chemical_list: D005609:Free Radicals; D045322:Photosynthetic Reaction Center Complex Proteins; D011809:Quinones