FIGURE 2.

Detection of a complex between CcmE, heme, and cytochrome c-b₅₆₂. A, Western blot of cell membranes using a cytochrome b₅₆₂ antibody. B, Western blot of cell membranes using a CcmE antibody. C, SDS-PAGE of cell membranes stained for proteins containing covalently bound heme. For each panel, the lane order is: M, molecular mass markers (as indicated, in kDa); lanes 1, cells expressing the Ccm system from pEC86; 2, cells expressing cytochrome c-b₅₆₂ R98C/Y101C; 3, cells expressing the Ccm system from pEC86 and cytochrome c-b₅₆₂ R98C/Y101C; 4, cells expressing the Ccm system from pEC86 and cytochrome c-b₅₆₂ R98C; 5, cells expressing the Ccm system from pEC86 and cytochrome c-b₅₆₂ Y101C. The position of the CcmE-heme-cytochrome c-b₅₆₂ complex, which runs just above a cytochrome c-b₅₆₂ dimer and just below the CcmE dimer, is indicated. An equal amount of total protein was loaded in each lane.