Photosynthesis Research

, Volume 132, Issue 3, pp 305–309 | Cite as

A simple and efficient method to prepare pure dimers and monomers of the cytochrome b6f complex from spinach

  • María A. Luján
  • Patricia Lorente
  • Valter Zazubovich
  • Rafael Picorel
Technical Communication

Abstract

Using a single size-exclusion chromatography we were able to isolate highly pure dimers and monomers of the Cyt b6f 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 b6f 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 b6f 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.

Keywords

Cytochrome b6f Photosynthesis Plant Protein Purification SDS-PAGE 

Notes

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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Copyright information

© Springer Science+Business Media Dordrecht 2017

Authors and Affiliations

  1. 1.CSIC-Estación Experimental de Aula DeiZaragozaSpain
  2. 2.Department of PhysicsConcordia UniversityMontrealCanada

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