title: Mutational control of bioenergetics of bacterial reaction center probed by delayed fluorescence.
abstract: The free energy gap between the metastable charge separated state P(+)QA(-) and the excited bacteriochlorophyll dimer P* was measured by delayed fluorescence of the dimer in mutant reaction center proteins of the photosynthetic bacterium Rhodobacter sphaeroides. The mutations were engineered both at the donor (L131L, M160L, M197F and M202H) and acceptor (M265I and M234E) sides. While the donor side mutations changed systematically the number of H-bonds to P, the acceptor side mutations modified the energetics of QA by altering the van-der-Waals and electronic interactions (M265IT) and H-bond network to the acidic cluster around QB (M234EH, M234EL, M234EA and M234ER). All mutants decreased the free energy gap of the wild type RC (~890meV), i.e. destabilized the P(+)QA(-) charge pair by 60-110meV at pH8. Multiple modifications in the hydrogen bonding pattern to P resulted in systematic changes of the free energy gap. The destabilization showed no pH-dependence (M234 mutants) or slight increase (WT, donor-side mutants and M265IT above pH8) with average slope of 10-15meV/pH unit over the 6-10.5pH range. In wild type and donor-side mutants, the free energy change of the charge separation consisted of mainly enthalpic term but the acceptor side mutants showed increased entropic (even above that of enthalpic) contributions. This could include softening the structure of the iron ligand (M234EH) and the QA binding pocket (M265IT) and/or increase of the multiplicity of the electron transfer of charge separation in the acceptor side upon mutation.
authors.affiliation: Laboratoire de Chimie Physique UMR 8000, Batiment 350, Orsay-Cedex, Université de Paris-Sud, 91405, France.
authors.forename: Delphine
authors.initials: D
authors.lastname: Onidas
authors.forename: Gábor
authors.initials: G
authors.lastname: Sipka
authors.forename: Emese
authors.initials: E
authors.lastname: Asztalos
authors.forename: Péter
authors.initials: P
authors.lastname: Maróti
mesh_terms: D001665:Binding Sites; D004579:Electron Transport; D004734:Energy Metabolism; D005453:Fluorescence; D006860:Hydrogen Bonding; D007700:Kinetics; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D050505:Mutant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009154:Mutation / Q000235:genetics; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012242:Rhodobacter sphaeroides / Q000235:genetics / Q000378:metabolism*; D013816:Thermodynamics
chemical_list: D050505:Mutant Proteins; D045322:Photosynthetic Reaction Center Complex Proteins
keywords: BChl; BPhe; Bacterial photosynthesis; Charge recombination; DL; H-bond network; LDAO; Light-induced free energy changes; N,N′-dimethyldodecylamine N-oxide; P, primary electron donor; Q(A) and Q(B); RC; Rba; Reaction center protein; Rhodobacter; Triton X-100; UQ; WT; a non-covalently linked bacteriochlorophyll dimer; bacteriopheophytin; delayed light; monomeric bacteriochlorophyll; octylphenol polyethylene glycol ether; primary and secondary quinones; reaction center; ubiquinone; wild type