Dynamics of chromophore binding to Lhc proteins in vivo and in vitro during operation of the xanthophyll cycle.
Three plant xanthophylls are components of the xanthophyll cycle in which, upon exposure of leaves to high light, the enzyme violaxanthin de-epoxidase (VDE) transforms violaxanthin into zeaxanthin via the intermediate antheraxanthin. Previous work () showed that xanthophylls are bound to Lhc proteins and that substitution of violaxanthin with zeaxanthin induces conformational changes and fluorescence quenching by thermal dissipation. We have analyzed the efficiency of different Lhc proteins to exchange violaxanthin with zeaxanthin both in vivo and in vitro. Light stress of Zea mays leaves activates VDE, and the newly formed zeaxanthin is found primarily in CP26 and CP24, whereas other Lhc proteins show a lower exchange capacity. The de-epoxidation system has been reconstituted in vitro by using recombinant Lhc proteins, recombinant VDE, and monogalactosyl diacylglycerol (MGDG) to determine the intrinsic capacity for violaxanthin-to-zeaxanthin exchange of individual Lhc gene products. Again, CP26 was the most efficient in xanthophyll exchange. Biochemical and spectroscopic analysis of individual Lhc proteins after de-epoxidation in vitro showed that xanthophyll exchange occurs at the L2-binding site. Xanthophyll exchange depends on low pH, implying that access to the binding site is controlled by a conformational change via lumenal pH. These findings suggest that the xanthophyll cycle participates in a signal transduction system acting in the modulation of light harvesting versus thermal dissipation in the antenna system of higher plants.
Dipartimento Scientifico e Tecnologico, Università di Verona, Strada Le Grazie, 37134 Verona, Italy.
Tomas
T
Morosinotto
Roberta
R
Baronio
Roberto
R
Bassi
D002734:Chlorophyll / Q000378:metabolism; D000077194:Chlorophyll A; D004851:Epoxide Hydrolases / Q000378:metabolism; D005989:Glycerides / Q000378:metabolism; D008027:Light; D045342:Light-Harvesting Protein Complexes; D010084:Oxidation-Reduction; D045322:Photosynthetic Reaction Center Complex Proteins / Q000378:metabolism*; D018515:Plant Leaves / Q000378:metabolism; D018087:Plastids / Q000378:metabolism*; D013053:Spectrophotometry; D024341:Xanthophylls / Q000378:metabolism*; D003313:Zea mays / Q000378:metabolism*; D019207:beta Carotene / Q000031:analogs & derivatives* / Q000378:metabolism
D005989:Glycerides; D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins; D024341:Xanthophylls; D019207:beta Carotene; D002734:Chlorophyll; C005613:violaxanthin; C037184:chlorophyll b; D004851:Epoxide Hydrolases; D000077194:Chlorophyll A