The photosynthetic cytochrome c 550 from the diatom Phaeodactylum tricornutum
The photosynthetic cytochrome c 550 from the marine diatom Phaeodactylum tricornutum has been purified and characterized. Cytochrome c 550 is mostly obtained from the soluble cell extract in relatively large amounts. In addition, the protein appeared to be truncated in the last hydrophobic residues of the C-terminus, both in the soluble cytochrome c 550 and in the protein extracted from the membrane fraction, as deduced by mass spectrometry analysis and the comparison with the gene sequence. Interestingly, it has been described that the C-terminus of cytochrome c 550 forms a hydrophobic finger involved in the interaction with photosystem II in cyanobacteria. Cytochrome c 550 was almost absent in solubilized photosystem II complex samples, in contrast with the PsbO and Psb31 extrinsic subunits, thus suggesting a lower affinity of cytochrome c 550 for the photosystem II complex. Under iron-limiting conditions the amount of cytochrome c 550 decreases up to about 45% as compared to iron-replete cells, pointing to an iron-regulated synthesis. Oxidized cytochrome c 550 has been characterized using continuous wave EPR and pulse techniques, including HYSCORE, and the obtained results have been interpreted in terms of the electrostatic charge distribution in the surroundings of the heme centre.
KeywordsCytochrome c550 Phaeodactylum Photosystem II EPR Hemeprotein
Cytochrome c 550
Cytochrome c 6
Electron paramagnetic resonance
Hyperfine sublevel correlation spectroscopy
- MALDI-TOF MS
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
The authors thank Rocío Rodríguez (Proteomic Service, IBVF) for technical assistance, and Prof. Tatsuya Tomo (Tokyo University of Science, Japan) for the Psb31 antibodies.
This work was supported by the Spanish Ministry of Economy and Competitiveness (BIO2012-35271, BIO2015-64169-P, MAT2011-23861 and CTQ2015-64486-R) the Andalusian Government (PAIDI BIO-022) and the Aragón Government (Grupo consolidado B-18). All these grants were partially financed by the EU FEDER Program.
- Ago H, Adachi H, Umena Y, Tashiro T, Kawakami K, Kamiya N, Tian L, Han G, Kuang T, Liu Z, Wang F, Zou H, Enami I, Miyano M, Shen J-R (2016) Novel features of eukaryotic photosystem II revealed by its crystal structure analysis from a red alga. J Biol Chem 291:5676–5687CrossRefPubMedPubMedCentralGoogle Scholar
- Enami I, Iwai M, Akiyama A, Suzuki T, Okumura A, Katoh T, Tada O, Ohta H, Shen J-R (2003) Comparison of binding and functional properties of two extrinsic components, Cyt c550 and a 12 kDa protein, in cyanobacterial PSII with those in red algal PSII. Plant Cell Physiol 44:820–827CrossRefPubMedGoogle Scholar
- Kerfeld CA, Sawaya MR, Bottin H, Tran KT, Sugiura M, Cascio D, Desbois A, Yeates TO, Kirilovsky D, Boussac A (2003) Structural and EPR characterization of the soluble form of cytochrome c-550 and of the psbV2 gene product from the cyanobacterium Thermosynechococcus elongatus. Plant Cell Physiol 44:697–706CrossRefPubMedGoogle Scholar
- Martínez-Fábregas J, Díaz-Moreno I, González-Arzola K, Janocha S, Navarro JA, Hervás M, Bernhardt R, Velázquez-Campoy A, Díaz-Quintana A, De la Rosa MA (2014) Structural and functional analysis of novel human cytochrome c targets in apoptosis. Mol Cell Proteomics 13:1439–1456CrossRefPubMedPubMedCentralGoogle Scholar
- Molina-Heredia FP, Hervás M, Navarro JA, De la Rosa MA (1998) Cloning and correct expression in Escherichia coli of the petE and petJ genes respectively encoding plastocyanin and cytochrome c 6 from the cyanobacterium Anabaena sp. PCC 7119. Biochem Biophys Res Commun 243:302–306CrossRefPubMedGoogle Scholar
- Moore JK, Doney SC, Glover DM, Fung IY (2002) Iron cycling and nutrient-limitation patterns in surface waters of the world ocean. Deep Sea Res II(49):463–507Google Scholar
- Nagao R, Ishii A, Tada O, Suzuki T, Dohmae N, Okumura A, Iwai M, Takahashi T, Kashino Y, Enami I (2007) Isolation and characterization of oxygen-evolving thylakoid membranes and Photosystem II particles from a marine diatom Chaetoceros gracilis. Biochim Biophys Acta 1767:1353–1362CrossRefPubMedGoogle Scholar
- Nagao R, Moriguchi A, Tomo T, Niikura A, Nakajima S, Suzuki T, Okumura A, Iwai M, Shen J-R, Ikeuchi M, Enami I (2010a) Binding and functional properties of five extrinsic proteins in oxygen-evolving photosystem II from a marine centric diatom, Chaetoceros gracilis. J Biol Chem 285:29191–29199Google Scholar
- Nagao R, Tomo T, Noguchi E, Nakajima S, Suzuki T, Okumura A, Kashino Y, Mimuro M, Ikeuchi M, Enami I (2010b) Purification and characterization of a stable oxygen-evolving Photosystem II complex from a marine centric diatom, Chaetoceros gracilis. Biochim Biophys Acta 1797:160–166Google Scholar
- Navarro JA, Hervás M, De la Rosa MA (2011) Purification of plastocyanin and cytochrome c6 from plants, green algae and cyanobacteria. In: Carpentier R (ed) Photosynthesis protocols, vol 684. Humana Press, Totowa, pp 79–94Google Scholar
- Okumura A, Nagao R, Suzuki T, Yamagoe S, Iwai M, Nakazato K, Enami I (2008) A novel protein in Photosystem II of a diatom Chaetoceros gracilis is one of the extrinsic proteins located on lumenal side and directly associates with PSII core components. Biochim Biophys Acta 1777:1545–1551CrossRefPubMedGoogle Scholar
- Roncel M, González-Rodríguez AA, Naranjo B, Bernal-Bayard P, Lindahl AM, Hervás M, Navarro JA, Ortega JM (2016) Iron deficiency induces a partial inhibition of the photosynthetic electron transport and a high sensitivity to light in the diatom Phaeodactylum tricornutum. Front Plant Sci 7:1050CrossRefPubMedPubMedCentralGoogle Scholar
- Schweiger A, Jeschke G. (2001) Principles of pulse electron paramagnetic resonance. Oxford University Press, OxfordGoogle Scholar