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. 2015 May 22;290(28):17116–17130. doi: 10.1074/jbc.M115.659300

FIGURE 7.

FIGURE 7.

Binding specificity of Xyn10C-XBD and its isolated domains. a, representative binding of different carbohydrates to Xyn10C-XBD by ITC. The ligands in the syringe (20 mm oligosaccharides, 4 mg/ml polysaccharide) were titrated into Xyn10C-XBD loaded in the cell (46–60 μm). Measurements were performed at 25 °C in 50 mm sodium hydrogen/dihydrogen phosphate, pH 7. The upper panels show the raw data upon injection of 1, 5 × 5, 8 × 10, and 8 × 20 μl for xylotetraose; 1, 5 × 2, 5 × 3, 5 × 5, 4 × 10, and 9 × 20 μl for xylopentaose; and 1, 15 × 5, 4 × 10, and 21 × 20 μl for oat spelt arabinoxylan. The bottom panels show the integrated areas (symbols) obtained from respective raw data and their theoretical fit (continuous lines) using the two-sets of sites binding-model of the Origin ITC software with the best fitting thermodynamic parameters summarized in Table 3. b, nondenaturing polyacrylamide gels containing no ligand (control) or soluble xylans. Lane 1, BSA; lane 2, Xyn10C-XBD; lane 3, CBM22-1; lane 4, CBM22-2.