title: Bacterial photosynthetic reaction centers in trehalose glasses: coupling between protein conformational dynamics and electron-transfer kinetics as studied by laser-flash and high-field EPR spectroscopies.
abstract: The coupling between electron transfer (ET) and the conformational dynamics of the cofactor−protein complex in photosynthetic reaction centers (RCs) from Rhodobacter sphaeroides in water/glycerol solutions or embedded in dehydrated poly(vinyl alcohol) (PVA) films or trehalose glasses is reported. Matrix effects were studied by time-resolved 95 GHz high-field electron paramagnetic resonance (EPR) spectroscopy at room (290 K) and low (150 K) temperature. ET from the photoreduced quinone acceptor (QA•−) to the photo-oxidized donor (P865•+) is strongly matrix-dependent at room temperature: In the trehalose glasses, the recombination kinetics of P865•+QA•−, probed by EPR and optical spectroscopies, is faster and broadly distributed as compared to that of RCs in solution, reflecting the inhibition of the RC relaxation from the dark- to the light-adapted conformational substate and the hindrance of substate interconversion. Similarly accelerated kinetics was observed also in PVA at a water-to-RC molar ratio 10-fold lower than in trehalose. Despite the matrix dependence of the ET kinetics, continuous-wave (cw) EPR and electron spin echo (ESE) analyses of the photogenerated P865•+ and QA•− radical ions and P865•+QA•− radical pairs do not reveal significant matrix effects, at either 290 or 150 K, indicating no change in the molecular radical-pair configuration of the P865•+ and QA•− cofactors. Furthermore, the field dependences of the transverse relaxation times T2 of QA•− essentially coincide in trehalose and PVA at 290 K. T2 is similar in these two matrixes and in the glycerol/water system at 150 K, implying that the librational dynamics of QA•− are also unaffected by the matrix. We infer that the relative geometry of the primary donor and acceptor, as well as the local dynamics and hydrogen bonding of QA in its binding pocket, are not involved in the stabilization of P865•+QA•−. We suggest that the RC relaxation occurs rather by changes throughout the protein/solvent system. The control of the RC dynamics and ET by the environment is discussed, particularly with respect to the extraordinary efficacy of trehalose matrixes in restricting the RC motional degrees of freedom at elevated temperatures.
authors.affiliation: Max-Planck-Institut für Bioanorganische Chemie, 45470 Mülheim an der Ruhr, Germany.
authors.forename: Anton
authors.initials: A
authors.lastname: Savitsky
authors.forename: Marco
authors.initials: M
authors.lastname: Malferrari
authors.forename: Francesco
authors.initials: F
authors.lastname: Francia
authors.forename: Giovanni
authors.initials: G
authors.lastname: Venturoli
authors.forename: Klaus
authors.initials: K
authors.lastname: Möbius
mesh_terms: D004578:Electron Spin Resonance Spectroscopy; D004579:Electron Transport; D006860:Hydrogen Bonding; D007700:Kinetics; D007834:Lasers; D056004:Molecular Dynamics Simulation; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry*; D011487:Protein Conformation; D012242:Rhodobacter sphaeroides / Q000201:enzymology; D014199:Trehalose / Q000737:chemistry*
chemical_list: D045322:Photosynthetic Reaction Center Complex Proteins; D014199:Trehalose