title: A simple and efficient method to prepare pure dimers and monomers of the cytochrome b6f complex from spinach.
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.
authors.affiliation: CSIC-Estación Experimental de Aula Dei, Avda. Montañana 1005, 50059, Zaragoza, Spain.
authors.forename: María A
authors.initials: MA
authors.lastname: Luján
authors.affiliation: CSIC-Estación Experimental de Aula Dei, Avda. Montañana 1005, 50059, Zaragoza, Spain.
authors.forename: Patricia
authors.initials: P
authors.lastname: Lorente
authors.affiliation: Department of Physics, Concordia University, 7141 Sherbrook Str. West, Montreal, QC, H4B 1R6, Canada.
authors.forename: Valter
authors.initials: V
authors.lastname: Zazubovich
authors.affiliation: CSIC-Estación Experimental de Aula Dei, Avda. Montañana 1005, 50059, Zaragoza, Spain. picorel@eead.csic.es.
authors.forename: Rafael
authors.identifier: http://orcid.org/0000-0003-3791-129X
authors.initials: R
authors.lastname: Picorel
mesh_terms: D002736:Chloroplasts / Q000378:metabolism; D003573:Cytochrome b Group / Q000378:metabolism; D045346:Cytochrome b6f Complex / Q000378:metabolism; D003580:Cytochromes / Q000378:metabolism; D045303:Cytochromes b / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D018724:Spinacia oleracea / Q000378:metabolism*
chemical_list: D003573:Cytochrome b Group; D003580:Cytochromes; D045303:Cytochromes b; D045346:Cytochrome b6f Complex
keywords: Cytochrome b 6 f; Photosynthesis; Plant; Protein; Purification; SDS-PAGE