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Orange Carotenoid Protein Burrows Into the Phycobilisome to Provide Photoprotection

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Orange Carotenoid Protein Burrows Into the Phycobilisome to Provide Photoprotection Orange carotenoid protein burrows into the phycobilisome to provide photoprotection. Dvir Harris, Ofir Tal, Denis Jallet, Adjélé Wilson, Diana Kirilovsky, Noam Adir To cite this version: Dvir Harris, Ofir Tal, Denis Jallet, Adjélé Wilson, Diana Kirilovsky, et al.. Orange carotenoid protein burrows into the phycobilisome to provide photoprotection.. Proceedings of the National Academy of Sciences of the United States of America , National Academy of Sciences, 2016, 113 (12), pp.E1655-62. 10.1073/pnas.1523680113. hal-01457540 HAL Id: hal-01457540 https://hal.archives-ouvertes.fr/hal-01457540 Submitted on 27 May 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Distributed under a Creative Commons Attribution - NonCommercial| 4.0 International License Orange carotenoid protein burrows into the PNAS PLUS phycobilisome to provide photoprotection Dvir Harrisa, Ofir Tala, Denis Jalletb,c,1, Adjélé Wilsonb,c, Diana Kirilovskyb,c,2, and Noam Adira,2 aSchulich Faculty of Chemistry, Technion, Haifa 32000, Israel; bInstitute for Integrative Biology of the Cell (I2BC), Commissariat à l’Energie Atomique (CEA), CNRS, Université Paris-Sud, Université Paris-Saclay, 91198 Gif-sur-Yvette, France; and cCEA, Institut de Biologie et Technologies de Saclay (iBiTec-S), 91191 Gif-sur-Yvette, France Edited by Robert Haselkorn, University of Chicago, Chicago, IL, and approved February 3, 2016 (received for review December 4, 2015) In cyanobacteria, photoprotection from overexcitation of photochem- the C-terminal domain (residues 196–317; Cterm) is composed of ical centers can be obtained by excitation energy dissipation at the β-strands in a twisted sheet. The first 19 N-terminal amino acids (the level of the phycobilisome (PBS), the cyanobacterial antenna, induced “cap”)extendoverandinteractwitha solvent-exposed face of the by the orange carotenoid protein (OCP). A single photoactivated OCP β-sheet of the Cterm. The keto group of hECN is involved in hy- bound to the core of the PBS affords almost total energy dissipation. drogen bonding with Tyr201 and Trp288 in the Cterm (13, 16, 18). The precise mechanism of OCP energy dissipation is yet to be fully It was found that the Nterm containing the carotenoid is sufficient determined, and one question is how the carotenoid can approach any to induce quenching activity without the presence of the Cterm core phycocyanobilin chromophore at a distance that can promote (19, 20). Very recently, the structure of the N-terminal domain efficient energy quenching. We have performed intersubunit of the activated state of the OCPR (denoted RCP; red carot- cross-linking using glutaraldehyde of the OCP and PBS followed enoid protein) was determined to high resolution, revealing that by liquid chromatography coupled to tandem mass spectrometry upon release of the Cterm the carotenoid molecule translates (LC/MS-MS) to identify cross-linked residues. The only residues of 12 Å further into the Nterm (20). By this movement, hECN is the OCP that cross-link with the PBS are situated in the linker region, protected from the solvent, and this movement is accompanied between the N- and C-terminal domains and a single C-terminal res- by conformational changes that result in the change in carot- idue. These links have enabled us to construct a model of the site of enoid absorption. It can be inferred from these results that upon OCP binding that differs from previous models. We suggest that light activation the Cterm and Nterm of the OCP disassemble, BIOPHYSICS AND the N-terminal domain of the OCP burrows tightly into the PBS while allowing the Nterm to tightly associate with the PBS and perform COMPUTATIONAL BIOLOGY leaving the OCP C-terminal domain on the exterior of the complex. its functional role of energy transfer disruption. Circular dichroism Further analysis shows that the position of the small core linker protein (CD) spectra of the two isolated domains suggest that each set of ApcC is shifted within the cylinder cavity, serving to stabilize the secondary structures is not altered by their physical separation interaction between the OCP and the PBS. This is confirmed by a ΔApcC (19), as confirmed by the RCP crystal structure for the Nterm. The mutant. Penetration of the N-terminal domain can bring the OCP ca- length that links the Nterm to the Cterm is at most 37 residues rotenoidtowithin5–10 Å of core chromophores; however, alteration (160–196), and thus the two domains can reach a separation of of the core structure may be the actual source of energy dissipation. 60–80 Å, potentially leaving the Cterm unattached to the PBS (20). In the OCPo form the Cterm prevents the OCP from binding photosynthesis | light harvesting | nonphotochemical quenching | to the PBS, and in the OCPR the Cterm interacts with the fluo- cyanobacteria | cross-linking rescence recovery protein (FRP), an additional component of the NPQ system. Presence of the FRP greatly enhances the reversal of R O xcess energy arriving at photochemical reaction centers (RCs) OCP to OCP and unbinding of the OCP from the PBS (21). Ecan be detrimental (1, 2), leading to loss of photosynthetic vi- ability (photoinhibition; PI) (3, 4). Cyanobacteria have evolved Significance several mechanisms for energy dissipation to deal with over- excitation (5, 6). The major light-harvesting complex (LHC) Protection from overexcitation is one of the most important in cyanobacteria is the phycobilisome (PBS), a giant complex that requirements of all photosynthetic organisms. Here we present can functionally transfer energy to two to four RCs (7–9). If over- a model based on coupled cross-linking/mass spectrometry and excitation occurs rapidly, photoprotection can be achieved by de- site-directed mutagenesis of the means by which the orange creasing the energy arriving at the RCs by increasing energy thermal carotenoid protein (OCP) binds to the phycobilisome (PBS) an- dissipation (nonphotochemical quenching; NPQ) or by physical (or tenna complex to avoid photodamage. The model shows that functional) disconnection of the PBS (10, 11). In most cyanobacterial the protein must actively burrow into the complex, separating species, NPQ is obtained by the presence a 35-kDa, water-soluble, the PBS rings in the process. This penetration explains for the orange carotenoid protein (OCP) (12, 13). OCP-dependent NPQ first time, to our knowledge, how the OCP carotenoid could was shown to be induced (14, 15) by strong blue-green light. The approach the PBS chromophores at a distance that enables OCP has two states—an inactive resting orange state (OCPO)and nonphotochemical quenching. However, the alteration in the an active red state (OCPR). Upon illumination, the OCP un- core structure caused by OCP binding could also prevent en- dergoes carotenoid and protein conformational changes to yield ergy transmission to the reaction centers. the metastable OCPR (16). The OCPR binds to the PBS and sig- Author contributions: D.H., D.K., and N.A. designed research; D.H., D.J., and A.W. performed nificantly quenches excitation energy and PBS fluorescence (17). research; O.T. contributed new reagents/analytic tools; D.H., O.T., D.J., A.W., D.K., and N.A. The OCP noncovalently binds a single keto-carotenoid chromo- analyzed data; and D.H., D.K., and N.A. wrote the paper. ′ phore, 3 -hydroxyechinenone (hECN). The high-resolution crystal The authors declare no conflict of interest. O Synechocystis structure of the OCP [from the cyanobacterium sp. This article is a PNAS Direct Submission. PCC 6803; Syn; Protein Data Bank (PDB) ID code 3MG1] shows 1Present address: Department of Biology, Colorado State University, Fort Collins, CO 80523. a two-domain structure connected by an unstructured loop, and 2To whom correspondence may be addressed. Email: [email protected] or nadir@tx. revealed hECN to be concealed deep inside the protein scaffold technion.ac.il. and stabilized via hydrogen bonds from both domains (17). The This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10. N-terminal domain (residues 15–160; Nterm) is all α-helical, and 1073/pnas.1523680113/-/DCSupplemental. www.pnas.org/cgi/doi/10.1073/pnas.1523680113 PNAS Early Edition | 1of8 Fig. 1. Partial model of the Syn PBS. (A) The PBS is visualized from the direction of the thylakoid membrane. The model includes a single-basal core cylinder (including ApcA, ApcB, ApcC, ApcD, ApcE, and ApcF; colored in yellow, blue, light orange, gray, light green, and light blue, respectively) and a three-hexamer rod that includes the phycobiliproteins CpcA and CpcB (colored in pink and red, respectively) and the CpcC, CpcD, and CpcG linkers (in black). In all schematic models, linkers are shown as small objects because their position and structures are not known. Only the phycobiliprotein-type domain of ApcE is shown. (B) The same segment of the PBS is shown perpendicular to that in A (along the membrane). Colors are as in A. The core of the PBS is made up of two to five cylinders (Fig. S1), In the study presented here, we have analyzed the interactions R each composed of four (αβ)3 trimers of allophycocyanin (APC) between the OCP and the PBS using the cross-linking mass (22, 23). Two basal core cylinders sit on top of the membrane, and spectrometry (MS) methodology mentioned above with the more contain the main form of APC (APC660) and one copy each of three general linker glutaraldehyde (GA).
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