Effect of uncoupler on assembly pathway for pigment-binding protein of bacterial photosynthetic membranes.

The uncoupler carbonylcyanide m-chlorophenylhydrazone (CCCP) was used to investigate membrane protein assembly in the phototrophic bacterium Rhodobacter capsulatus. As found for Escherichia coli (T. Date, G. Zwizinsky, S. Ludmerer, and W. Wickner, Proc. Natl. Acad. Sci. 77:827-831, 1980) and mitochondrial proteins (N. Nelson and G. Schatz, Proc. Natl. Acad. Sci. USA 76:4365-4369, 1979), assembly across the bacterial photosynthetic membranes was sensitive to CCCP. At uncoupler concentrations which were sufficient to block the export of the periplasmic cytochrome c2 and an outer membrane protein, the integration of pigment-binding protein into the photosynthetic apparatus was abolished. The unassembled protein was detected on the inner surface of the intracytoplasmic membrane. After inactivation of CCCP, accumulated protein continued insertion into the membrane. The data suggest that after binding to the cytoplasmic face of the membrane, translocation of protein into a transmembrane orientation takes place, which is a prerequisite for the formation of a functional pigment-protein complex.