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A simple and efficient method to prepare pure dimers and monomers of the cytochrome b 6 f complex from spinach

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Abstract

Using a single size-exclusion chromatography we were able to isolate highly pure dimers and monomers of the Cyt b 6 f complex from spinach from a bulk preparation of that protein complex obtained with a standard procedure. At higher protein/detergent ratio during the chromatography most of the Cyt b 6 f complex remained as dimers. In contrast, at lower protein/detergent ratio (around 15 times lower), most dimers became monomerized. As a bonus, this chromatography also allowed the elimination of potential Chl a contaminant to the Cyt b 6 f preparations. SDS-PAGE protein analysis with 18% (w/v) acrylamide revealed the loss of the ISP subunit in our monomeric preparation. However, it fully retained the content of Chl a, a prerequisite to perform any spectroscopic study involving this unique pigment.

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Acknowledgements

This work was supported by the Ministry of Economy and Competiveness of Spain (Grant No. AGL2014-55300-R) and Aragon Government (Grant E33).

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Authors and Affiliations

  1. CSIC-Estación Experimental de Aula Dei, Avda. Montañana 1005, 50059, Zaragoza, Spain

    María A. Luján, Patricia Lorente & Rafael Picorel

  2. Department of Physics, Concordia University, 7141 Sherbrook Str. West, Montreal, QC, H4B 1R6, Canada

    Valter Zazubovich

Authors
  1. María A. Luján
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  2. Patricia Lorente
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  3. Valter Zazubovich
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  4. Rafael Picorel
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Correspondence to Rafael Picorel.

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Luján, M.A., Lorente, P., Zazubovich, V. et al. A simple and efficient method to prepare pure dimers and monomers of the cytochrome b 6 f complex from spinach. Photosynth Res 132, 305–309 (2017). https://doi.org/10.1007/s11120-017-0375-x

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  • DOI: https://doi.org/10.1007/s11120-017-0375-x

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