FIGURE 11.
Thermal stability of wild type, F119L, or R141H phosphomannomutase2 monitored by thermal shift assay. The profiles (A, WT; B, F119L; C, R141H) were assessed by thermal shift assays. Proteins (0.275 mg/ml in 20 mm Hepes, pH 7.5, 2 mm MgCl2, 150 mm NaCl, 1 mm dithiothreitol, 2.4× SYPRO Orange) were heated from 25 to 80 °C at 0.5 °C/min, and fluorescence was recorded (excitation, 490 nm; emission, 575 nm). The experiment was performed in the presence of no ligand (light blue), 5 mm EDTA (blue), 0.5 mm Glc-1-P (red), 0.5 mm Glc-1-P plus 0.5 mm vanadate (black), and 0.5 mm Glc-1,6-P2 (green).