title: Essential role of two tyrosines and two tryptophans on the photoprotection activity of the Orange Carotenoid Protein.
abstract: Photosynthetic organisms have developed photoprotective mechanisms to protect themselves from lethal high light intensities. One of these mechanisms involves the dissipation of excess absorbed light energy into heat. In cyanobacteria, light activation of a soluble carotenoid protein, the Orange Carotenoid Protein (OCP), binding a keto carotenoid, is the key inducer of this mechanism. Blue-green light absorption triggers structural changes within the carotenoid and the protein, leading to the conversion of a dark orange form into a red active form. Here we report the role in photoconversion and photoprotection of individual conserved tyrosines and tryptophans surrounding the rings of the carotenoid. Our results demonstrate that the interaction between the keto group of the carotenoid and Tyr201 and Trp288 is essential for OCP photoactivity. In addition, these amino acids are responsible for carotenoid affinity and specificity. We have already demonstrated that the aromatic character of Tyr44 and Trp110 interacting with the hydroxyl ring is critical. Here we show that the replacement of Tyr44 by Ser affects the stability of the red form avoiding its accumulation at any temperature, while Trp110Ser is affected in the energy necessary to the orange to red conversion and in the interaction with the antenna. Collectively our data support the idea that the red form is essential for photoprotection but not sufficient. Specific conformational changes occurring in the protein seem to be critical to the events leading to energy dissipation.
authors.affiliation: Commissariat à l'Energie Atomique (CEA), Institut de Biologie et Technologies de Saclay (iBiTec-S), France.
authors.forename: Adjélé
authors.initials: A
authors.lastname: Wilson
authors.forename: Claire
authors.initials: C
authors.lastname: Punginelli
authors.forename: Mohea
authors.initials: M
authors.lastname: Couturier
authors.forename: François
authors.initials: F
authors.lastname: Perreau
authors.forename: Diana
authors.initials: D
authors.lastname: Kirilovsky
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D002338:Carotenoids / Q000378:metabolism; D000458:Cyanobacteria / Q000235:genetics / Q000378:metabolism*; D006860:Hydrogen Bonding; D008027:Light; D010777:Photochemistry; D010788:Photosynthesis / Q000528:radiation effects*; D011487:Protein Conformation; D014364:Tryptophan / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; C109473:orange carotenoid protein, Synechocystis; D002338:Carotenoids; D014443:Tyrosine; D014364:Tryptophan