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. 2009 Jun 29;185(7):1195–1207. doi: 10.1083/jcb.200812025

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© 2009 Saint-Marcoux et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 5. — Coimmunoprecipitation analysis of CCBs and cyt b₆ interactions. Digitonin-solubilized membranes (prepared as for BN-PAGE) from the petB-C35V strain were incubated with protein A–Sepharose beads coupled to antibodies against peptides of CCB3 (α-CCB3), subunit IV N terminus (α-IV), cyt b₆ C terminus (α-b₆C), and cyt b₆ N terminus (α-b₆N), or against respective preimmune serum (pi). (a) Coimmunoprecipitations with α-CCB3 and α-IV. (b) Coimmunoprecipitations with α-b₆C and α-b₆N. Aliquots of digitonin-solubilized membranes (sol) and immunoprecipitates (IP) were separated by SDS-PAGE and analyzed by immunoblotting. (c) Top view from the n side of the membrane and (d) side view in the membrane plane of the cyt b₆ structure. Images in c and d were generated using PyMol and the coordinates from pdb file 1Q90 with α-b₆N peptide antigen in red, α-b₆C peptide antigen in green, b_L and b_H-hemes in brown, c_i-heme in orange, and Cys35 in yellow.