Skip to main content
. Author manuscript; available in PMC: 2019 Apr 23.
Published in final edited form as: Biochemistry. 2016 Oct 12;55(42):5917–5926. doi: 10.1021/acs.biochem.6b00826

Fig. 3. α-NPG binding in the absence or presence of a bound Nb.

Fig. 3.

α-NPG at 1 mM or 0.2 mM was injected into the sample cell containing WT LacY (100 μM) or G46W/G262W mutant (28 μM), respectively (a, e). WT LacY or the mutant pre-incubated with Nb-9036 (b, f) or Nb-9065 (c, g) at a ratio of 1:1.75 or 2.5, respectively, was titrated with α-NPG (0.2 mM). DMSO (0.5%) was present in both titrand and titrant for α-NPG solubility. Thermogram (a-c, e-g) was recorded at 25 °C. Injection of α-NPG into buffer at a concentration used for the titration into the protein samples is shown in cyan (a, e), and injection of 1 mM nitrophenyl-α-glucoside into the WT LacY is shown in magenta (a). ΔQ (on the right side) vs α-NPG:LacY molar ratio (on the top) were fitted to a one-site independent binding model (blue curves), and the results were presented in Table 2 and panels d and f. (d, h) Comparison of α-NPG binding energy in the absence (black) and presence of Nb-9036 (cyan) or Nb-9065 (gray). Unpaired t-test was applied for the statastic anlaysis as deacribed in the legand to figure 1.