Skip to main content
. 2012 Sep 21;7(9):e45024. doi: 10.1371/journal.pone.0045024

Figure 2. Disrupting the hydrophobic interactions via mutating Leu46 alters the structural stability of MIF.

Figure 2

(A) Leu46 hydrophobic pocket of wt huMIF. VMD representations of the hydrophobic pocket, where Leu46 is mutated to a phenylalanine (L46F) (B), alanine (L46A) (C), or glycine (L46G) (D). (E-H): The three Leu46 mutants are structurally less stable than the wild type protein, but retain the same overall secondary structure. (E) Far-UV CD spectra of wt and Leu46 mutants. (F) Thermal denaturation of wt and Leu46 mutants (at 20 µM) followed by far-UV CD at 218 nm. (G) GdnHCl denaturation studies monitored by far-UV CD at 218 nm and fluorescence spectroscopy (H), excitation wavelength: 295 nm, protein concentration: 10 µM. All spectroscopic experiments were performed in PBS 1X, pH 7.4 buffer. Black lines, wt MIF; blue lines, L46F MIF; red lines, L46A MIF; green lines: L46G MIF.