Interactions between the Photosystem II Subunit PsbS and Xanthophylls Studied in Vivo and in Vitro

• Published in: The Journal of biological chemistry Vol. 283; no. 13; pp. 8434 - 8445
• Main Authors: Bonente, Giulia, Howes, Barry D., Caffarri, Stefano, Smulevich, Giulietta, Bassi, Roberto
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 28.03.2008; American Society for Biochemistry and Molecular Biology
• Subjects: Arabidopsis - enzymology; Arabidopsis - genetics; Circular Dichroism; Dicyclohexylcarbodiimide - metabolism; Genotype; Metabolism and Bioenergetics; Mutation - genetics; Phenotype; Photosynthetic Reaction Center Complex Proteins - chemistry; Photosynthetic Reaction Center Complex Proteins - genetics; Photosynthetic Reaction Center Complex Proteins - metabolism; Photosystem II Protein Complex - chemistry; Photosystem II Protein Complex - genetics; Photosystem II Protein Complex - metabolism; Plant Proteins - chemistry; Plant Proteins - genetics; Plant Proteins - metabolism; Protein Binding; Protein Folding; Protein Subunits - chemistry; Protein Subunits - genetics; Protein Subunits - metabolism; Spectrum Analysis, Raman; Xanthophylls - metabolism
• DOI: 10.1074/jbc.M708291200
• ISSN: 0021-9258

The photosystem II subunit PsbS is essential for excess energy dissipation (qE); however, both lutein and zeaxanthin are needed for its full activation. Based on previous work, two models can be proposed in which PsbS is either 1) the gene product where the quenching activity is located or 2) a proton-sensing trigger that activates the quencher molecules. The first hypothesis requires xanthophyll binding to two PsbS-binding sites, each activated by the protonation of a dicyclohexylcarbodiimide-binding lumen-exposed glutamic acid residue. To assess the existence and properties of these xanthophyll-binding sites, PsbS point mutants on each of the two Glu residues PsbS E122Q and PsbS E226Q were crossed with the npq1/npq4 and lut2/npq4 mutants lacking zeaxanthin and lutein, respectively. Double mutants E122Q/npq1 and E226Q/npq1 had no qE, whereas E122Q/lut2 and E226Q/lut2 showed a strong qE reduction with respect to both lut2 and single glutamate mutants. These findings exclude a specific interaction between lutein or zeaxanthin and a dicyclohexylcarbodiimide-binding site and suggest that the dependence of nonphotochemical quenching on xanthophyll composition is not due to pigment binding to PsbS. To verify, in vitro, the capacity of xanthophylls to bind PsbS, we have produced recombinant PsbS refolded with purified pigments and shown that Raman signals, previously attributed to PsbS-zeaxanthin interactions, are in fact due to xanthophyll aggregation. We conclude that the xanthophyll dependence of qE is not due to PsbS but to other pigment-binding proteins, probably of the Lhcb type.