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. Author manuscript; available in PMC: 2012 Nov 1.
Published in final edited form as: Mol Microbiol. 2011 Oct 10;82(3):634–647. doi: 10.1111/j.1365-2958.2011.07843.x

Fig. 5.

Fig. 5

Determination of the site of BMH cross-linking. (A) The AtxA protein, which contains six cysteine residues, has five putative motifs: a winged helix-turn-helix (WH), a Mga-like helix-turn-helix (HTH), two phosphotransferase regulation domains (PRD1 and PRD2), and an Enzyme IIB-like motif (EIIB). (B) Following induction with 30 μM IPTG, cells from cultures of atxA-null strains (UT376) containing pUTE658-derived atxA point mutants were lysed and treated with BMH. SDS-PAGE (4-15%) and Western blots with AtxA-specific antibody were used to detect various forms of AtxA. (C) A similar experiment was performed using derivatives of the parent strain (UT375) expressing atxA from its native locus and pUTE992-derived FLAG-tagged versions of AtxA or the AtxA point mutants. Westerns were probed with α-FLAG and α-AtxA antibodies. EV = empty vector. Molecular weights of the protein standards are as indicated (SeeBlue Plus2, Invitrogen).