FIG 3

FIG 3

PecA is subjected to self-cleavage. (A) Expression of PecA_myc and PecA_D293Gmyc in M. marinumΔpecA was detected by immunoblotting of whole-cell lysate. (B) Subcellular localization and secretion analysis of M. marinum wild type (indicated with “-”) or wild type expressing PecA_myc or PecA_D293Gmyc. WCL, whole-cell lysate; GP, Genapol-treated pellet (membrane associated and intracellular); GS, Genapol supernatant (membrane associated); CF, culture filtrate. GroEL2 was probed as a cytosolic control. (C) Immunoblot analysis of PecA_myc and PecA_D293Gmyc expressed in E. coli (whole-cell lysate). (D) Immunoblot analysis of in vitro translation of PecA_myc and PecA_D293Gmyc. (E) Immunoblot analysis of M. marinum wild type; wild type expressing PecA_myc, PecA_A84Dmyc, or PecA_AA88EDmyc; and ΔpecA mutant expressing PecA_D293Gmyc (whole-cell lysate). The arrow indicates the fully cleaved form of PecA. Proteins were detected by their C-terminal Myc tag with an anti-Myc antibody.