title: Functional consequences of the organization of the photosynthetic apparatus in Rhodobacter sphaeroides: II. A study of PufX- membranes.
abstract: In the bacterium R. sphaeroides, the polypeptide PufX is indispensable for photosynthetic growth. Its deletion is known to have important consequences on the organization of the photosynthetic apparatus. In the wild-type strain, complexes between the reaction center (RC) and the antenna (light-harvesting complex 1 (LH1)) are associated in dimers, and LH1 does not fully encircle the RC. In the absence of PufX, the complexes become monomeric, and the LH1 ring closes around the RC. We analyzed the functional consequences of PufX deletion. Some effects can be ascribed to the monomerization of the RC.LH1 complexes: the number of RCs that share a common antenna for excitation transfer or a common quinone pool become smaller. We examined the kinetic effects of the closed LH1 ring on quinone turnover: diffusion across LH1 entails a delay of approximately 1 ms, and the barrier appears to be located directly against the quinone-binding (secondary quinone acceptor (Q(B))) pocket. The diffusion of ubiquinol from the RC to the cytochrome bc1 complex is approximately 2-fold slower in the mutant, suggesting an increased distance between the two complexes. The properties of the Q(B) pocket (binding of inhibitors, stabilization of Q(B-), and rate of Q(B)-H2 formation) appear to be modified in the mutant. Another specificity of PufX- is the accumulation of closed centers in the Q(A-) (where Q(A) is the primary quinone acceptor) state as the secondary acceptor pool becomes reduced, which is probably the origin of photosynthetic incompetence. We suggest that this is related to the Q(B) pocket alterations. The malfunction of the reaction center is probably due to a faulty association with LH1 that is prevented in the PufX-containing structure.
authors.affiliation: Unité Mixte de Recherche 6191 CNRS-Commissariat à l'Energie Atomique-Aix Marseille II, Département d'Ecophysiologie Végétale et de Microbiologie, Commissariat à l'Energie Atomique Cadarache, 13108 Saint Paul-lez-Durance, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Comayras
authors.forename: Colette
authors.initials: C
authors.lastname: Jungas
authors.forename: Jérôme
authors.initials: J
authors.lastname: Lavergne
mesh_terms: D001426:Bacterial Proteins / Q000502:physiology*; D016227:Benzoquinones / Q000378:metabolism; D004058:Diffusion; D014450:Electron Transport Complex III / Q000378:metabolism; D045342:Light-Harvesting Protein Complexes / Q000378:metabolism / Q000502:physiology*; D010084:Oxidation-Reduction; D010788:Photosynthesis; D045322:Photosynthetic Reaction Center Complex Proteins / Q000378:metabolism; D012242:Rhodobacter sphaeroides / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D016227:Benzoquinones; D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins; C088749:PufX protein, Rhodobacter; C004532:quinone; D014450:Electron Transport Complex III