title: Removal and reconstitution of the carotenoid antenna of xanthorhodopsin.
abstract: Salinixanthin, a C(40)-carotenoid acyl glycoside, serves as a light-harvesting antenna in the retinal-based proton pump xanthorhodopsin of Salinibacter ruber. In the crystallographic structure of this protein, the conjugated chain of salinixanthin is located at the protein-lipid boundary and interacts with residues of helices E and F. Its ring, with a 4-keto group, is rotated relative to the plane of the π-system of the carotenoid polyene chain and immobilized in a binding site near the β-ionone retinal ring. We show here that the carotenoid can be removed by oxidation with ammonium persulfate, with little effect on the other chromophore, retinal. The characteristic CD bands attributed to bound salinixanthin are now absent. The kinetics of the photocycle is only slightly perturbed, showing a 1.5-fold decrease in the overall turnover rate. The carotenoid-free protein can be reconstituted with salinixanthin extracted from the cell membrane of S. ruber. Reconstitution is accompanied by restoration of the characteristic vibronic structure of the absorption spectrum of the antenna carotenoid, its chirality, and the excited-state energy transfer to the retinal. Minor modification of salinixanthin, by reducing the carbonyl C=O double bond in the ring to a C-OH, suppresses its binding to the protein and eliminates the antenna function. This indicates that the presence of the 4-keto group is critical for carotenoid binding and efficient energy transfer.
authors.affiliation: Department of Physiology and Biophysics, University of California, Irvine, CA 92697-4560, USA.
authors.forename: Eleonora S
authors.initials: ES
authors.lastname: Imasheva
authors.forename: Sergei P
authors.initials: SP
authors.lastname: Balashov
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Wang
authors.forename: Janos K
authors.initials: JK
authors.lastname: Lanyi
mesh_terms: D000645:Ammonium Sulfate / Q000737:chemistry; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D002338:Carotenoids / Q000737:chemistry* / Q000378:metabolism; D004735:Energy Transfer; D006027:Glycosides / Q000378:metabolism; D007700:Kinetics; D010084:Oxidation-Reduction; D025604:Rhodopsins, Microbial / Q000737:chemistry* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D006027:Glycosides; D025604:Rhodopsins, Microbial; C470489:salinixanthin; C519152:xanthorhodopsin, Salinibacter; C031276:ammonium peroxydisulfate; D002338:Carotenoids; D000645:Ammonium Sulfate