Different Functions of the Paralogs to the N-Terminal Domain of the Orange Carotenoid Protein in the Cyanobacterium <i>Anabaena</i> sp. PCC 7120

López‐Igual, Rocío; Wilson, Adjélé; Leverenz, Ryan L.; Melnicki, Matthew R.; Carbon, Céline Bourcier de; Sutter, Markus; Turmo, Aiko; Perreau, François; Kerfeld, Cheryl A.; Kirilovsky, Diana

doi:10.1104/pp.16.00502

Cited by 65 publications

(75 citation statements)

References 56 publications

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“…While the physiological relevance of the horizontal carotenoid transfer discovered here is unclear, yet, the possibility of carotenoid translocation from OCP-CTD to AnaCTDH strongly suggested high affinity of the latter to carotenoids and supported the idea that carotenoid transfer can occur in many directions, involving both NTDÁCTD(H) and CTDÁCTD(H) interactions. In line with this, it was found recently that AnaCTDH can retrieve carotenoid from AnaHCP1 (but not HCP2, HCP3, nor HCP4 [40]) suggesting that HCP1 may serve as an additional carotenoid carrier [44]. Therefore, we questioned whether the apparently high affinity of AnaCTDH may allow it to retrieve the carotenoid from other OCP-related proteins containing the NTD.…”

Section: Inverse Transfer Of Carotenoid To Ctdhsupporting

confidence: 53%

Light‐controlled carotenoid transfer between water‐soluble proteins related to cyanobacterial photoprotection

et al. 2019

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Carotenoids are lipophilic pigments with multiple biological functions from coloration to vision and photoprotection. Still, the number of water‐soluble carotenoid‐binding proteins described to date is limited, and carotenoid transport and carotenoprotein maturation processes are largely underexplored. Recent studies revealed that CTDHs, which are natural homologs of the C‐terminal domain (CTD) of the orange carotenoid protein (OCP), a photoswitch involved in cyanobacterial photoprotection, are able to bind carotenoids, with absorption shifted far into the red region of the spectrum. Despite the recent discovery of their participation in carotenoid transfer processes, the functional roles of the diverse family of CTDHs are not well understood. Here, we characterized CTDH carotenoproteins from Anabaena variabilis (AnaCTDH) and Thermosynechococcus elongatus and examined their ability to participate in carotenoid transfer processes with a set of OCP‐derived proteins. This revealed that carotenoid transfer occurs in several directions guided by different affinities for carotenoid and specific protein–protein interactions. We show that CTDHs have higher carotenoid affinity compared to the CTD of OCP from Synechocystis, which results in carotenoid translocation from the CTD into CTDH via a metastable heterodimer intermediate. Activation of OCP by light, or mutagenesis compromising the OCP structure, provides AnaCTDH with an opportunity to extract carotenoid from the full‐length OCP, either from Synechocystis or Anabaena. These previously unknown reactions between water‐soluble carotenoproteins demonstrate multidirectionality of carotenoid transfer, allowing for efficient and reversible control over the carotenoid‐mediated protein oligomerization by light, which gives insights into the physiological regulation of OCP activity by CTDH and suggests multiple applications.

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Section: Inverse Transfer Of Carotenoid To Ctdhsupporting

confidence: 53%

Light‐controlled carotenoid transfer between water‐soluble proteins related to cyanobacterial photoprotection

et al. 2019

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“…PCC 7120 have been described . The construction of plasmids pCDF-RCP-Syn-1-165Ctag, pCDF-NTD-ana-1-165Ctag, and pCDF-HCP4-4941Ctag was described previously López-Igual et al, 2016).…”

Section: Construction Of Ocp Ntd and Hcp Plasmids For Expression Inmentioning

confidence: 99%

“…The crtBEIY operon in pACBETA was expressed constitutively under the control of the crtE promoter, whereas the crtW gene was under the control of the arabinose-inducible promoter araBAD and the ocp, ctdh, and hcp genes were under the control of a T7 RNA polymerase promoter, and their expression was enhanced by the addition of isopropylthio-b-galactoside. The expression of different genes and the isolation of apo-and holo-CTDHs and HCPs were described previously López-Igual et al, 2016). restriction site of pPSBA2.…”

Section: Pcdf-ctd6803-ctagmentioning

confidence: 99%

Paralogs of the C-Terminal Domain of the Cyanobacterial Orange Carotenoid Protein Are Carotenoid Donors to Helical Carotenoid Proteins

et al. 2017

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The photoactive Orange Carotenoid Protein (OCP) photoprotects cyanobacteria cells by quenching singlet oxygen and excess excitation energy. Its N-terminal domain is the active part of the protein, and the C-terminal domain regulates the activity. Recently, the characteristics of a family of soluble carotenoid-binding proteins (Helical Carotenoid Proteins [HCPs]), paralogs of the N-terminal domain of OCP, were described. Bioinformatics studies also revealed the existence of genes coding for homologs of CTD. Here, we show that the latter genes encode carotenoid proteins (CTDHs). This family of proteins contains two subgroups with distinct characteristics. One CTDH of each clade was further characterized, and they proved to be very good singlet oxygen quenchers. When synthesized in Escherichia coli or Synechocystis PCC 6803, CTDHs formed dimers that share a carotenoid molecule and are able to transfer their carotenoid to apo-HCPs and apo-OCP. The CTDHs from clade 2 have a cysteine in position 103. A disulfide bond is easily formed between the monomers of the dimer preventing carotenoid transfer. This suggests that the transfer of the carotenoid could be redox regulated in clade 2 CTDH. We also demonstrate here that apoOCPs and apo-CTDHs are able to take the carotenoid directly from membranes, while HCPs are unable to do so. HCPs need the presence of CTDH to become holo-proteins. We propose that, in cyanobacteria, the CTDHs are carotenoid donors to HCPs.Photosynthetic organisms performing oxygenic photosynthesis use solar energy, water, and inorganic carbon to produce all the organic molecules they need.Photosynthesis converts the absorbed energy into chemical energy and the reduction power necessary for the assimilation of CO 2 and the synthesis of organic carbon molecules. However, photosynthetic organisms cannot control the incoming flux of light, and too much light generates secondary reactions creating dangerous species of oxygen leading to cellular damage. Thus, in order to survive, photosynthetic organisms have developed a large variety of photoprotective mechanisms. One of them decreases the energy arriving at the reaction centers by increasing the thermal dissipation of excess excitation energy at the level of the antennae (for review, see Niyogi and Truong, 2013). In cyanobacteria, a soluble carotenoid protein, the Orange Carotenoid Protein (OCP), is essential for this mechanism, known as the OCP-related nonphotochemical quenching mechanism (Wilson et al., 2006; for review, see Kirilovsky and Kerfeld, 2016). In addition, OCP has a second photoprotective activity. It is a very good singlet oxygen ( 1 O 2 ) quencher (Kerfeld et al., 2003;Sedoud et al., 2014).OCP is a photoactive soluble carotenoid protein (Wilson et al., 2008). It is composed of two globular domains: an a-helical N-terminal domain (NTD; residues 18-165) that is unique to cyanobacteria and an a-helix/b-sheet C-terminal domain (CTD; residues

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“…Moreover, upon mixing of COCP(CAN) with the apoprotein form of the NTD (Apo-RCP), spectral and functional features of an OCP O -like state appeared, which readily underwent photoconversion resulting in the formation of RCP that was capable of PBs fluorescence quenching [25]. In effect, these findings revealed an as yet unprecedented carotenoid transfer mechanism initiated by COCP suggesting that isolated COCP homologs in cyanobacteria may play some role in carotenoid mobilization and storage, besides acting as singlet oxygen quenchers [23,25,26]. The modular architecture of OCP and the possibility to assemble it in fully functional form from its separated domains allows tailoring the use of carotenoid-binding entities according to highly specialized functions needed by the cell.…”

Section: Introductionmentioning

confidence: 97%

The Unique Protein-To-Protein Carotenoid Transfer Mechanism

Maksimov

Sluchanko

Slonimskiy

et al. 2017

Preprint

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Orange Carotenoid Protein (OCP) is known to be an effector and regulator of cyanobacterial photoprotection. This 35 kDa water-soluble protein provides specific environment for keto-carotenoids, the excitation of which induced by the absorption of bluegreen light causes dramatic but fully reversible rearrangements of the OCP structure, including carotenoid translocation and separation of C-and N-terminal domains upon transition from the basic orange to photoactivated red OCP form. While recent studies significantly improved our understanding of the OCP photocycle and interaction with phycobilisomes and the fluorescence not peer-reviewed) is the author/funder. All rights reserved. No reuse allowed without permission.The copyright holder for this preprint (which was . http://dx.doi.org/10.1101/127183 doi: bioRxiv preprint first posted online Apr. 13, 2017; 2 recovery protein, the mechanism of OCP assembly remains unclear. Apparently, this process requires targeted delivery and incorporation of a highly hydrophobic carotenoid molecule into the water-soluble apoprotein of OCP. Recently, we introduced a novel carotenoid carrier protein, COCP, which consists of dimerized C-domain(s) of OCP and can combine with the isolated Ndomain to form transient OCP-like species. Here, we demonstrate that in vitro COCP efficiently transfers otherwise tightly bound carotenoid to the full-length OCP apoprotein, resulting in formation of the photoactive OCP from completely photoinactive species. We accurately analyze peculiarities of this carotenoid transfer process which, to the best of our knowledge, seems unique, previously uncharacterized protein-to-protein carotenoid transfer process. We hypothesize that a similar OCP assembly can occur in vivo, substantiating specific roles of the COCP carotenoid carrier in cyanobacterial photoprotection.

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Different Functions of the Paralogs to the N-Terminal Domain of the Orange Carotenoid Protein in the Cyanobacterium Anabaena sp. PCC 7120

Cited by 65 publications

References 56 publications

Light‐controlled carotenoid transfer between water‐soluble proteins related to cyanobacterial photoprotection

Light‐controlled carotenoid transfer between water‐soluble proteins related to cyanobacterial photoprotection

Paralogs of the C-Terminal Domain of the Cyanobacterial Orange Carotenoid Protein Are Carotenoid Donors to Helical Carotenoid Proteins

The Unique Protein-To-Protein Carotenoid Transfer Mechanism

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