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. Author manuscript; available in PMC: 2012 Jun 3.
Published in final edited form as: J Mol Biol. 2011 Apr 1;409(2):146–158. doi: 10.1016/j.jmb.2011.03.027

Figure 2.

Figure 2

Identification of the epitopic region of TcpF recognized by mAb13. (A) Diagram of peptides. Seven TcpF peptides were constructed, each with an N-terminal His6x tag and a C-terminal intein tag. (B) mAb13 recognizes Fpep4, Fpep5, and Fpep7. Immunoblots of whole cell lysate of V. cholerae or peptide-expressing E. coli strains were first blotted with mAb13 then stripped and re-probed with the TcpF polyclonal antisera and an anti-His antibody to evaluate peptide stability and antibody reactivity. WT indicates culture supernatant from V. cholerae O395 grown under TCP inducing conditions. The 36 kDa TcpF band is recognized by both TcpF antibodies. The calculated molecular weights of the full-length peptides, including the His6x and intein tags, are as follows: Fpep1, 65.2 kDa; Fpep2, 73.2; Fpep3, 66.5; Fpep4, 67.9 Fpep5, 66.0; Fpep6, 83.8; Fpep7, 78.0. (C) Amino acid sequence of the TcpF epitopic region, which corresponds to the sequence shared by Fpep4, Fpep5 and Fpep7. Critical binding residues Glu251 and Glu252 are underlined.