1 CSIC-Estación Experimental de Aula Dei, Avda. Montañana 1005, 50059, Zaragoza, Spain.
2 Department of Physics, Concordia University, 7141 Sherbrook Str. West, Montreal, QC, H4B 1R6, Canada.
3 CSIC-Estación Experimental de Aula Dei, Avda. Montañana 1005, 50059, Zaragoza, Spain. picorel@eead.csic.es.

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

Photosynth Res. 2017 Jun;132(3):305-309

Authors: Luján MA, Lorente P, Zazubovich V, Picorel R

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.

PMID: 28374305 [PubMed - indexed for MEDLINE]