FIG. 6.
CcoH binds to a preassembled cbb3-Cox. (A) In vitro-synthesized wild-type CcoH (wt) or the truncated CcoH versions CcoH-M0 and CcoH-M1 were in vitro synthesized in the presence of the indicated membranes. MT1131 contains wild-type amounts of cbb3-Cox, while in GK32, cbb3-Cox is deleted. M7G corresponds to an R. capsulatus ccoP mutant that assembles the 210-kDa CcoNO subcomplex. After in vitro synthesis, samples were separated by BN-PAGE, and radioactively labeled bands were detected by phosphorimaging. Asterisks (*) indicate the fully assembled 230-kDa cbb3-Cox complex, while plus signs (+) indicate the 210-kDa subcomplex lacking the CcoP subunit. (B) Synthesis control of the in vitro reactions described for panel A. Note that the three panels correspond to the same gel, which was split into three parts. Therefore, the size differences among the three CcoH derivatives are not visible.