The protease-protected SecA 65-kDa domain lies near the N terminus of the protein. The [35S]SecA 65-kDa fragments resulting from limited trypsin treatment of SecA in solution (S, lanes 1, 3, 5, and 7) or protected from tryptic digestion under translocation conditions (TL, lanes 2, 4, 6, and 8) were analyzed by peptide mapping using S. aureus V8, as described. The black arrow identifies the position of the SecA 65-kDa domain.