Skip to main content
. Author manuscript; available in PMC: 2017 Mar 17.
Published in final edited form as: J Mol Biol. 2007 May 10;370(4):701–713. doi: 10.1016/j.jmb.2007.04.075

Figure 5.

Figure 5

Analysis of binding of PspA1 mutant to LF. Various PspA1 residues (Glu180, Glu182, Asp237 and Glu279) involved in binding to NLF were mutated to alanine. The resulting mutant protein was expressed and used in binding studies. After incubation of PspA1 (wild-type and mutant) with LF (NLF and rhLF), the mixtures were subjected to gel electrophoresis on a non-denaturing gel. For Western blot analysis proteins were then transferred to a nitrocellulose membrane and detected using monoclonal mouse IgG Xi126 (as primary antibody) and alkaline phosphate-conjugated anti-mouse IgG antibody (as secondary ones). (a) Non-denaturing gel. (b) Western blot. 1, PspA1 (residues 1–288); 2, PspA1+ NLF; 3, PspA1+ rhLF; 4, PspA1 mutant; 5, PspA1 mutant+ NLF; 6, PspA1 mutant+ rhLF; 7, NLF; 8, rhLF.