title: The Ile(L229) → Met mutation impairs the quinone binding to the QB-pocket in reaction centers of Rhodobacter sphaeroides.
abstract: A spontaneous mutant (R/89) of photosynthetic purple bacterium Rhodobacter sphaeroides R-26 was selected for resistance to 200 μM atrazin. It showed increased resistance to interquinone electron transfer inhibitors of o-phenanthroline (resistance factor, RF=20) in UQo reconstituted isolated reaction centers and terbutryne in reaction centers (RF=55) and in chromatophores (RF=85). The amino acid sequence of the QB binding protein of the photosynthetic reaction center (the L subunit) was determined by sequencing the corresponding pufL gene and a single mutation was found (Ile(L229) → Met). The changed amino acid of the mutant strain is in van der Waals contact with the secondary quinone QB. The binding and redox properties of QB in the mutant were characterized by kinetic (charge recombination) and multiple turnover (cytochrome oxidation and semiquinone oscillation) assays of the reaction center. The free energy for stabilization of QAQB (-) with respect to QA (-)QB was ΔGAB=-60 meV and 0 meV in reaction centers and ΔGAB=-85 meV and -46 meV in chromatophores of R-26 and R/89 strains at pH 8, respectively. The dissociation constants of the quinone UQo and semiquinone UQo (-) in reaction centers from R-26 and R/89 showed significant and different pH dependence. The observed changes in binding and redox properties of quinones are interpreted in terms of differential effects (electrostatics and mesomerism) of mutation on the oxidized and reduced states of QB.
authors.affiliation: Department of Biophysics, József Attila University, Egyetem u.2., H-6722, Szeged, Hungary.
authors.forename: J
authors.initials: J
authors.lastname: Tandori
authors.forename: L
authors.initials: L
authors.lastname: Nagy
authors.forename: A
authors.initials: A
authors.lastname: Puskás
authors.forename: M
authors.initials: M
authors.lastname: Droppa
authors.forename: G
authors.initials: G
authors.lastname: Horváth
authors.forename: P
authors.initials: P
authors.lastname: Maróti