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A comparative study of three signaling forms of the orange carotenoid protein

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Abstract

Orange carotenoid protein (OCP) is a water-soluble photoactive protein responsible for a photoprotective mechanism of nonphotochemical quenching in cyanobacteria. Under blue–green illumination, OCP converts from the stable orange into the signaling red quenching form; however, the latter form could also be obtained by chemical activation with high concentrations of sodium thiocyanate (NaSCN) or point mutations. In this work, we show that a single replacement of tryptophan-288, normally involved in protein-chromophore interactions, by alanine, results in formation of a new protein form, hereinafter referred to as purple carotenoid protein (PCP). Comparison of resonance Raman spectra of the native photoactivated red form, chemically activated OCP, and PCP reveals that carotenoid conformation is sensitive to the structure of the C-domain, implicating that the chromophore retains some interactions with this part of the protein in the active red form. Combination of differential scanning fluorimetry and picosecond time-resolved fluorescence anisotropy measurements allowed us to compare the stability of different OCP forms and to estimate relative differences in protein rotation rates. These results were corroborated by hydrodynamic analysis of proteins by dynamic light scattering and analytical size-exclusion chromatography, indicating that the light-induced conversion of the protein is accompanied by a significant increase in its size. On the whole, our data support the idea that the red form of OCP is a molten globule-like protein in which, however, interactions between the carotenoid and the C-terminal domain are preserved.

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Abbreviations

OCPO :

Orange carotenoid protein—orange form

OCPR :

Orange carotenoid protein—red form

PCP:

Purple Carotenoid Protein (W288A-OCP)

RCP:

Red Carotenoid Protein

ECN:

Echinenone

NPQ:

Non-photochemical quenching

τ :

Fluorescence lifetime

φ fl :

Fluorescence quantum yield

r :

Anisotropy

θ :

Correlation time

NR:

Nile red

SEC:

Size-exclusion chromatography

DSF:

Differential scanning fluorimetry

DLS:

Dynamic light scattering

HOOP:

Hydrogen-out-of-plane

References

  • Bartalucci G, Coppin J, Fisher S, Hall G, Helliwell JR, Helliwell M, Liaaen-Jensen S (2007) Unravelling the chemical basis of the bathochromic shift in the lobster carapace; new crystal structures of unbound astaxanthin, canthaxanthin and zeaxanthin. Acta Crystallogr B 63:328–337. doi:10.1107/S0108768106052633

    Article  CAS  PubMed  Google Scholar 

  • Berera R, van Stokkum IH, Gwizdala M, Wilson A, Kirilovsky D, van Grondelle R (2012) The photophysics of the orange carotenoid protein, a light-powered molecular switch. J Phys Chem B 116:2568–2574. doi:10.1021/jp2108329

    Article  CAS  PubMed  Google Scholar 

  • Boulay C, Abasova L, Six C, Vass I, Kirilovsky D (2008) Occurrence and function of the orange carotenoid protein in photoprotective mechanisms in various cyanobacteria. Biochim Biophys Acta 1777:1344–1354. doi:10.1016/j.bbabio.2008.07.002

    Article  CAS  PubMed  Google Scholar 

  • Breitenbach J, Gerjets T, Sandmann G (2013) Catalytic properties and reaction mechanism of the CrtO carotenoid ketolase from the cyanobacterium Synechocystis sp. PCC 6803. Arch Biochem Biophys 529:86–91

    Article  CAS  PubMed  Google Scholar 

  • Chabera P, Durchan M, Shih PM, Kerfeld CA, Polivka T (2011) Excited-state properties of the 16 kDa red carotenoid protein from Arthrospira maxima. Biochim Biophys Acta 1807:30–35. doi:10.1016/j.bbabio.2010.08.013

    Article  CAS  PubMed  Google Scholar 

  • Chergui M et al (2013) Comparing the photophysics of the two forms of the Orange carotenoid protein using 2D electronic spectroscopy. EPJ Web of Conferences 41:08008. doi:10.1051/epjconf/20134108008

  • Chib R, Raut S, Sabnis S, Singhal P, Gryczynski Z, Gryczynski I (2014) Associated anisotropy decays of ethidium bromide interacting with DNA. Methods Appl Fluoresc 2(1):015003. doi:10.1088/2050-6120/2/1/015003

    Article  PubMed  PubMed Central  Google Scholar 

  • Cianci M, Rizkallah PJ, Olczak A, Raftery J, Chayen NE, Zagalsky PF, Helliwell JR (2002) The molecular basis of the coloration mechanism in lobster shell: beta-crustacyanin at 3.2-A resolution. Proc Natl Acad Sci USA 99:9795–9800. doi:10.1073/pnas.152088999

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • de Carbon CB, Thurotte A, Wilson A, Perreau F, Kirilovsky D (2015) Biosynthesis of soluble carotenoid holoproteins in Escherichia coli. Sci Rep 5:9085. doi:10.1038/srep09085

    Article  PubMed Central  Google Scholar 

  • Demmig-Adams B, Adams WW (1996) The role of xanthophyll cycle carotenoids in the protection of photosynthesis. Trends Plant Sci 1:21–26

    Article  Google Scholar 

  • Denison FC, Gökirmak T, Ferl RJ (2014) Phosphorylation-related modification at the dimer interface of 14-3-3ω dramatically alters monomer interaction dynamics. Arch Biochem Biophys 541:1–12

    Article  CAS  PubMed  Google Scholar 

  • Fernández-González B, Sandmann G, Vioque A (1997) A new type of asymmetrically acting β-carotene ketolase is required for the synthesis of echinenone in the cyanobacterium Synechocystis sp. PCC 6803. J Biol Chem 272:9728–9733

    Article  PubMed  Google Scholar 

  • Flora K, Brennan JD, Baker GA, Doody MA, Bright FV (1998) Unfolding of acrylodan-labeled human serum albumin probed by steady-state and time-resolved fluorescence methods. Biophys J 75:1084–1096

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Frank HA, Cogdell RJ (1996) Carotenoids in Photosynthesis. Photochem Photobiol 63:257–264. doi:10.1111/j.1751-1097.1996.tb03022.x

    Article  CAS  PubMed  Google Scholar 

  • Graf M, Bojak A, Deml L, Bieler K, Wolf H, Wagner R (2000) Concerted action of multiple cis-acting sequences is required for Rev dependence of late human immunodeficiency virus type 1 gene expression. J Virol 74:10822–10826

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gupta S et al (2015) Local and global structural drivers for the photoactivation of the orange carotenoid protein. Proc Natl Acad Sci USA 112:E5567–E5574. doi:10.1073/pnas.1512240112

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • King JD, Liu H, He G, Orf GS, Blankenship RE (2014) Chemical activation of the cyanobacterial orange carotenoid protein. FEBS Lett 588:4561–4565. doi:10.1016/j.febslet.2014.10.024

    Article  CAS  PubMed  Google Scholar 

  • Kirilovsky D, Kerfeld CA (2013) The Orange Carotenoid Protein: a blue-green light photoactive protein. Photochem Photobiol Sci 12:1135–1143. doi:10.1039/c3pp25406b

    Article  CAS  PubMed  Google Scholar 

  • Kish E, Pinto MM, Kirilovsky D, Spezia R, Robert B (2015) Echinenone vibrational properties: from solvents to the orange carotenoid protein. Biochim Biophys Acta 1847:1044–1054. doi:10.1016/j.bbabio.2015.05.010

    Article  CAS  PubMed  Google Scholar 

  • Lakowicz JR (2013) Principles of fluorescence spectroscopy. Springer, Berlin

    Google Scholar 

  • Leverenz RL, Jallet D, Li MD, Mathies RA, Kirilovsky D, Kerfeld CA (2014) Structural and functional modularity of the orange carotenoid protein: distinct roles for the N- and C-terminal domains in cyanobacterial photoprotection. Plant Cell 26:426–437. doi:10.1105/tpc.113.118588

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Leverenz RL et al (2015) A 12 Å carotenoid translocation in a photoswitch associated with cyanobacterial photoprotection. Science 348:3

    Article  Google Scholar 

  • Liu WL, Wang ZG, Zheng ZR, Li AH, Su WH (2008) Effect of β-ring rotation on the structures and vibrational spectra of β-carotene: density functional theory analysis. J Phys Chem A 112(42):10580–10585

    Article  CAS  PubMed  Google Scholar 

  • Liu H, Zhang H, Niedzwiedzki DM, Prado M, He G, Gross ML, Blankenship RE (2013) Phycobilisomes supply excitations to both photosystems in a megacomplex in cyanobacteria. Science 342:1104–1107

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Liu H, Zhang H, King JD, Wolf NR, Prado M, Gross ML, Blankenship RE (2014) Mass spectrometry footprinting reveals the structural rearrangements of cyanobacterial orange carotenoid protein upon light activation. Biochim Biophys Acta 1837:1955–1963. doi:10.1016/j.bbabio.2014.09.004

    Article  CAS  PubMed  Google Scholar 

  • Liu H et al (2016) Dramatic Domain Rearrangements of the Cyanobacterial Orange Carotenoid Protein upon Photoactivation. Biochemistry 55:1003–1009. doi:10.1021/acs.biochem.6b00013

    Article  CAS  PubMed  Google Scholar 

  • Maksimov EG et al (2014) The time course of non-photochemical quenching in phycobilisomes of Synechocystis sp. PCC6803 as revealed by picosecond time-resolved fluorimetry. Biochim Biophys Acta 1837:1540–1547. doi:10.1016/j.bbabio.2014.01.010

    Article  CAS  PubMed  Google Scholar 

  • Maksimov EG et al (2015) The Signaling State of Orange Carotenoid Protein. Biophys J 109:595–607. doi:10.1016/j.bpj.2015.06.052

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Misawa N et al (1995) Structure and functional analysis of a marine bacterial carotenoid biosynthesis gene cluster and astaxanthin biosynthetic pathway proposed at the gene level. J Bacteriol 177:6575–6584

    CAS  PubMed  PubMed Central  Google Scholar 

  • Niesen FH, Berglund H, Vedadi M (2007) The use of differential scanning fluorimetry to detect ligand interactions that promote protein stability. Nat Protoc 2(9):2212–2221. doi:10.1038/nprot.2007.321

    Article  CAS  PubMed  Google Scholar 

  • Pascal AA et al (2005) Molecular basis of photoprotection and control of photosynthetic light-harvesting. Nature 436:134–137

    Article  CAS  PubMed  Google Scholar 

  • Polívka T, Kerfeld CA, Pascher T, Sundström V (2005) Spectroscopic Properties of the Carotenoid 3‘-Hydroxyechinenone in the orange carotenoid protein from the Cyanobacterium Arthrospira maxima. Biochemistry 44:3994–4003. doi:10.1021/bi047473t

    Article  PubMed  Google Scholar 

  • Salares V, Young N, Bernstein H, Carey P (1979) Mechanisms of spectral shifts in lobster carotenoproteins The resonance raman spectra of ovoverdin and the crustacyanins. Biochim Biophys Acta (BBA) Protein Struct 576:176–191

    Article  CAS  Google Scholar 

  • Shakhnovich EI, Finkelstein AV (1989) Theory of cooperative transitions in protein molecules. I. Why denaturation of globular protein is a first-order phase transition. Biopolymers 28:1667–1680. doi:10.1002/bip.360281003

    Article  CAS  PubMed  Google Scholar 

  • Stadnichuk IN, Yanyushin MF, Maksimov EG, Lukashev EP, Zharmukhamedov SK, Elanskaya IV, Paschenko VZ (2012) Site of non-photochemical quenching of the phycobilisome by orange carotenoid protein in the cyanobacterium Synechocystis sp. PCC 6803. Biochim Biophys Acta 1817:1436–1445. doi:10.1016/j.bbabio.2012.03.023

    Article  CAS  PubMed  Google Scholar 

  • Stadnichuk IN et al (2013) Fluorescence quenching of the phycobilisome terminal emitter LCM from the cyanobacterium Synechocystis sp. PCC 6803 detected in vivo and in vitro. J Photochem Photobiol B 125:137–145. doi:10.1016/j.jphotobiol.2013.05.014

    Article  CAS  PubMed  Google Scholar 

  • Wilson A et al (2010) Structural determinants underlying photoprotection in the photoactive orange carotenoid protein of cyanobacteria. J Biol Chem 285:18364–18375. doi:10.1074/jbc.M110.115709

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wilson A, Punginelli C, Couturier M, Perreau F, Kirilovsky D (2011) Essential role of two tyrosines and two tryptophans on the photoprotection activity of the Orange Carotenoid Protein. Biochim Biophys Acta 1807:293–301. doi:10.1016/j.bbabio.2010.12.009

    Article  CAS  PubMed  Google Scholar 

  • Wilson A, Gwizdala M, Mezzetti A, Alexandre M, Kerfeld CA, Kirilovsky D (2012) The essential role of the N-terminal domain of the orange carotenoid protein in cyanobacterial photoprotection: importance of a positive charge for phycobilisome binding. Plant Cell 24:1972–1983. doi:10.1105/tpc.112.096909

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yguerabide J, Epstein HF, Stryer L (1970) Segmental flexibility in an antibody molecule. J Mol Biol 51:573–590

    Article  CAS  PubMed  Google Scholar 

  • Zhang H, Liu H, Niedzwiedzki DM, Prado M, Jiang J, Gross ML, Blankenship RE (2014) Molecular mechanism of photoactivation and structural location of the cyanobacterial orange carotenoid protein. Biochemistry 53:13–19. doi:10.1021/bi401539w

    Article  PubMed  Google Scholar 

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Acknowledgments

V.Z.P. and E.G.M. thank the Russian Foundation for Basic Research (Projects Nos. 14-04-01536A and 15-04-01930A) for partial support of this work. E.A.S. and E.G.M. thank the Russian Scientific Foundation (Project 14-17-00451) for partial support of this work. E.G.M., G.V.T., and K.E.K. thank the Russian Ministry of Education and Science (Project MK-5949.2015.4) for partial support of this work. This work was supported, in part by a Dynasty Foundation Fellowship to E.G.M. The reported study was funded by RFBR and Moscow city Government according to the research Project No. 15-34-70007 «mol_a_mos» . T.F., F.-J.S, N.N.T., M.W., and M.M gratefully acknowledge support from the Stifterverband für die Deutsche Wissenschaft (Study reform Project “OPLChem”) and the German Research Foundation (DFG, Cluster of Excellence “Unifying Concepts in Catalysis”).

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Authors and Affiliations

  1. Department of Biophysics, Faculty of Biology, M.V. Lomonosov Moscow State University, 119992, Moscow, Russia

    E. G. Maksimov, E. A. Parshina, K. E. Klementiev, G. V. Tsoraev, V. Z. Paschenko & A. B. Rubin

  2. Institute of Chemistry PC 14, Technical University of Berlin, Straße des 17. Juni 135, 10623, Berlin, Germany

    M. Moldenhauer, N. N. Tavraz, M. Willoweit, F.-J. Schmitt & T. Friedrich

  3. Department of Quantum Electronics, Faculty of Physics, M.V. Lomonosov Moscow State University, 119992, Moscow, Russia

    E. A. Shirshin

  4. A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, 119071, Moscow, Russia

    N. N. Sluchanko

  5. Institute for Molecular Biosciences, Johann Wolfgang Goethe University of Frankfurt, Max-von-Laue-Straße 9, 60438, Frankfurt, Germany

    J. Breitenbach & G. Sandmann

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  1. E. G. Maksimov
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  2. M. Moldenhauer
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Correspondence to E. G. Maksimov.

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Maksimov, E.G., Moldenhauer, M., Shirshin, E.A. et al. A comparative study of three signaling forms of the orange carotenoid protein. Photosynth Res 130, 389–401 (2016). https://doi.org/10.1007/s11120-016-0272-8

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  • DOI: https://doi.org/10.1007/s11120-016-0272-8

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