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. 2012 Aug 4;287(40):33796–33811. doi: 10.1074/jbc.M112.388579

FIGURE 4.

FIGURE 4.

NMR titrations of MBHA with α3,α6-mannopentaose. A, superposition of the two-dimensional 1H-15N HSQC spectra of free (black) and α3,α6-mannopentaose-bound MBHA at 1:6 molar ratio (magenta) of MBHA to α3,α6-mannopentaose. The four amide resonances of MBHA that correspond to Gly26 and Gly93 in OAA or PFA are displayed in the bottom inset. Three selected regions that are discussed under “Results” are indicated by dashed boxes. B–E, superposition of selected regions of the two-dimensional 1H-15N HSQC spectra of free (black) and α3,α6-mannopentaose bound MBHA at 1:1 (red), 1:2 (green), 1:4 (blue), and 1:6 (magenta) molar ratios of MBHA to sugar for residues Ala15/Ala82/Ala149/Ala216 (B), residues Tyr53/Tyr120/Tyr187/Tyr257 (C), residues Ser13/Thr80/Ser147/Ser214 (D), and residues Gly60/Gly127/Gly194/Gly261 (E). All spectra are plotted at the same contour level, and all titrations were carried out using 0.100 mm MBHA in 20 mm sodium acetate, 20 mm NaCl, 3 mm NaN3, 90/10% H2O/D2O (pH 5.0), 25 °C.