title: Preparative purification of the subunits of chloroplast and Rhodospirillum rubrum coupling factors by flat-bed electrofocusing in granulated gels.
abstract: A rapid method for the preparative purification of the subunits of oligomeric proteins like chloroplast and Rhodospirillum rubrum coupling factors is presented. It involves the dissociation of the protein in urea and the separation of its subunits by isoelectric focusing in flat-beds of Sepharose CL-4B or Sephadex G-75 superfine, in the presence of urea and in an overnight run. Using this procedure in the pH range 5-7, we have purified to homogeneity the alpha, beta and delta subunits of chloroplast coupling factor, as well as the alpha and beta subunits of Rhodospirillum rubrum coupling factor. The full separation of the gamma and epsilon subunits of chloroplast coupling factor, which focused at the same pH, was achieved by gel filtration high-performance liquid chromatography.
mesh_terms: D007525:Isoelectric Focusing / Q000379:methods; D046911:Macromolecular Substances; D006180:Proton-Translocating ATPases / Q000032:analysis*; D012247:Rhodospirillum rubrum / Q000201:enzymology*
chemical_list: D046911:Macromolecular Substances; D006180:Proton-Translocating ATPases