Abstract
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.
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Abbreviations
- ß-DM:
-
ß-dodecyl-maltoside
- Cc550 :
-
Cytochrome c 550
- Cc6 :
-
Cytochrome c 6
- CW:
-
Continuous wave
- EPR:
-
Electron paramagnetic resonance
- HYSCORE:
-
Hyperfine sublevel correlation spectroscopy
- MALDI-TOF MS:
-
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
- PSII:
-
Photosystem II
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Acknowledgements
The authors thank Rocío Rodríguez (Proteomic Service, IBVF) for technical assistance, and Prof. Tatsuya Tomo (Tokyo University of Science, Japan) for the Psb31 antibodies.
Funding
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.
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Bernal-Bayard, P., Puerto-Galán, L., Yruela, I. et al. The photosynthetic cytochrome c 550 from the diatom Phaeodactylum tricornutum . Photosynth Res 133, 273–287 (2017). https://doi.org/10.1007/s11120-016-0327-x
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DOI: https://doi.org/10.1007/s11120-016-0327-x