A simple and efficient method to prepare pure dimers and monomers of the cytochrome b 6 f complex from spinach
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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.
KeywordsCytochrome b6f Photosynthesis Plant Protein Purification SDS-PAGE
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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