title: The thylakoid proton motive force in vivo. Quantitative, non-invasive probes, energetics, and regulatory consequences of light-induced pmf.
abstract: Endogenous probes of light-induced transthylakoid proton motive force (pmf), membrane potential (Deltapsi) and DeltapH were used in vivo to assess in Arabidopsis the lumen pH responses of regulatory components of photosynthesis. The accumulation of zeaxanthin and protonation of PsbS were found to have similar pK(a) values, but quite distinct Hill coefficients, a feature allowing high antenna efficiency at low pmf and fine adjustment at higher pmf. The onset of "energy-dependent' exciton quenching (q(E)) occurred at higher lumen pH than slowing of plastoquinol oxidation at the cytochrome b(6)f complex, presumably to prevent buildup of reduced electron carriers that can lead to photodamage. Quantitative comparison of intrinsic probes with the electrochromic shift signal in situ allowed quantitative estimates of pmf and lumen pH. Within a degree of uncertainly of approximately 0.5 pH units, the lumen pH was estimated to range from approximately 7.5 (under weak light at ambient CO(2)) to approximately 5.7 (under 50 ppm CO(2) and saturating light), consistent with a 'moderate pH' model, allowing antenna regulation but preventing acid-induced photodamage. The apparent pK(a) values for accumulation of zeaxanthin and PsbS protonation were found to be approximately 6.8, with Hill coefficients of about 4 and 1 respectively. The apparent shift between in vitro violaxanthin deepoxidase protonation and zeaxanthin accumulation in vivo is explained by steady-state competition between zeaxanthin formation and its subsequent epoxidation by zeaxanthin epoxidase. In contrast to tobacco, Arabidopsis showed substantial variations in the fraction of pmf (0.1-0.7) stored as Deltapsi, allowing a more sensitive qE response, possible as an adaptation to life at lower light levels.
authors.affiliation: Institute of Biological Chemistry, Washington State University, Pullman, WA 99164-6340, USA.
authors.forename: Kenji
authors.initials: K
authors.lastname: Takizawa
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Cruz
authors.forename: Atsuko
authors.initials: A
authors.lastname: Kanazawa
authors.forename: David M
authors.initials: DM
authors.lastname: Kramer
mesh_terms: D017360:Arabidopsis / Q000378:metabolism; D002245:Carbon Dioxide / Q000378:metabolism; D045347:Cytochromes b6 / Q000378:metabolism; D045348:Cytochromes f / Q000378:metabolism; D004583:Electrons; D005456:Fluorescent Dyes; D007700:Kinetics; D008027:Light; D010084:Oxidation-Reduction; D010971:Plastoquinone / Q000031:analogs & derivatives / Q000378:metabolism; D018892:Proton-Motive Force; D020524:Thylakoids / Q000378:metabolism*
chemical_list: D045347:Cytochromes b6; D005456:Fluorescent Dyes; D002245:Carbon Dioxide; C003165:plastoquinol; D045348:Cytochromes f; D010971:Plastoquinone