title: The accumulation of the light-harvesting 2 complex during remodeling of the Rhodobacter sphaeroides intracytoplasmic membrane results in a slowing of the electron transfer turnover rate of photochemical reaction centers.
abstract: A functional proteomic analysis of the intracytoplasmic membrane (ICM) development process was performed in Rhodobacter sphaeroides during adaptation from high-intensity illumination to indirect diffuse light. This initiated an accelerated synthesis of the peripheral light-harvesting 2 (LH2) complex relative to that of LH1-reaction center (RC) core particles. After 11 days, ICM vesicles (chromatophores) and membrane invagination sites were isolated by rate-zone sedimentation and subjected to clear native gel electrophoresis. Proteomic analysis of gel bands containing the RC-LH1 and -LH2 complexes from digitonin-solubilized chromatophores revealed high levels of comigrating electron transfer enzymes, transport proteins, and membrane assembly factors relative to their equivalent gel bands from cells undergoing adaptation to direct low-level illumination. The GroEL chaperonin accounted for >65% of the spectral counts in the RC-LH1 band from membrane invagination sites, which together with the appearance of a universal stress protein suggested that the viability of these cells was challenged by light limitation. Functional aspects of the photosynthetic unit assembly process were monitored by near-IR fast repetition rate analysis of variable fluorescence arising from LH-bacteriochlorophyll a components. The quantum yield of the primary charge separation during the early stages of adaptation showed a gradual increase (variable/maximal fluorescence = 0.78-0.83 between 0 and 4 h), while the initial value of ~70 for the functional absorption cross section (σ) gradually increased to 130 over 4 days. These dramatic σ increases showed a direct relation to gradual slowing of the RC electron transport turnover rate (τ(QA)) from ~1.6 to 6.4 ms and an ~3-fold slowing of the rate of reoxidation of the ubiquinone pool. These slowed rates are not due to changes in UQ pool size, suggesting that the relation between increasing σ and τ(QA) reflects the imposition of constraints upon free diffusion of ubiquinone redox species between the RC and cytochrome bc(1) complex as the membrane bilayer becomes densely packed with LH2 rings.
authors.affiliation: Department of Molecular Biology and Biochemistry, Rutgers University, Busch Campus, Piscataway, New Jersey 08854-8082, USA.
authors.forename: Kamil
authors.initials: K
authors.lastname: Woronowicz
authors.forename: Daniel
authors.initials: D
authors.lastname: Sha
authors.forename: Raoul N
authors.initials: RN
authors.lastname: Frese
authors.forename: Robert A
authors.initials: RA
authors.lastname: Niederman
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism*; D004579:Electron Transport; D005453:Fluorescence; D045342:Light-Harvesting Protein Complexes / Q000737:chemistry* / Q000378:metabolism; D010084:Oxidation-Reduction; D012242:Rhodobacter sphaeroides / Q000737:chemistry / Q000254:growth & development / Q000378:metabolism*; D014451:Ubiquinone / Q000737:chemistry / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D045342:Light-Harvesting Protein Complexes; D014451:Ubiquinone