title: De-epoxidation of violaxanthin in light-harvesting complex I proteins.
abstract: The conversion of violaxanthin (Vx) to zeaxanthin (Zx) in the de-epoxidation reaction of the xanthophyll cycle plays an important role in the protection of chloroplasts against photooxidative damage. Vx is bound to the antenna proteins of both photosystems. In photosystem II, the formation of Zx is essential for the pH-dependent dissipation of excess light energy as heat. The function of Zx in photosystem I is still unclear. In this work we investigated the de-epoxidation characteristics of light-harvesting complex proteins of photosystem I (LHCI) under in vivo and in vitro conditions. Recombinant LHCI (Lhcal-4) proteins were reconstituted with Vx and lutein, and the convertibility of Vx was studied in an in vitro assay using partially purified Vx de-epoxidase isolated from spinach thylakoids. All four LHCI proteins exhibited unique de-epoxidation characteristics. An almost complete Vx conversion to Zx was observed only in Lhca3, whereas Zx formation in the other LHCI proteins decreased in the order Lhca4 > Lhca1 > Lhca2. Most likely, these differences in Vx de-epoxidation were related to the different accessibility of the respective carotenoid binding sites in the distinct antenna proteins. The results indicate that Vx bound to site V1 and N1 is easily accessible for de-epoxidation, whereas Vx bound to L2 is only partially and/or with the slower kinetics convertible to Zx. The de-epoxidation properties determined for the monomeric recombinant proteins were consistent with those obtained for isolated native LHCI-730 and LHCI-680 in the same in vitro assay and the de-epoxidation state found under in vivo conditions in native LHCIs.
authors.affiliation: Institut für Biochemie der Pflanzen, Heinrich Heine-Universität Düsseldorf, Germany.
authors.forename: Antje
authors.initials: A
authors.lastname: Wehner
authors.forename: Stefanie
authors.initials: S
authors.lastname: Storf
authors.forename: Peter
authors.initials: P
authors.lastname: Jahns
authors.forename: Volkmar H R
authors.initials: VH
authors.lastname: Schmid
mesh_terms: D001059:Apoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D002734:Chlorophyll / Q000737:chemistry; D004852:Epoxy Compounds / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D007700:Kinetics; D045342:Light-Harvesting Protein Complexes / Q000378:metabolism; D018551:Solanum lycopersicum / Q000737:chemistry / Q000254:growth & development / Q000378:metabolism; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D045331:Photosystem I Protein Complex / Q000378:metabolism; D010860:Pigments, Biological / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018724:Spinacia oleracea / Q000737:chemistry; D020524:Thylakoids / Q000737:chemistry; D024341:Xanthophylls; D019207:beta Carotene / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism*
chemical_list: D001059:Apoproteins; D004852:Epoxy Compounds; D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins; D045331:Photosystem I Protein Complex; D010860:Pigments, Biological; D011994:Recombinant Proteins; D024341:Xanthophylls; D019207:beta Carotene; D002734:Chlorophyll; C005613:violaxanthin