Fig. 5

Fig. 5

Thermodynamic and folding stability evaluation of two missense variants using NanoDSF. a NanoDSF melting temperatures for WT Mm IPT1-3 (orange) and Mm IPT1-3 p.(Gly129Ser) variant (pink). Measurements show at least three T_m peaks (orange dotted line) for the WT IPT1-3, likely because the protein fragment includes multiple IPT domains that unfold sequentially. Measured T_m values are shifted to the left (pink dotted line) showing a decrease of the thermal-folding stability. Temperatures are labeled for each T_m transition point. b Results for WT Mm IPT5-6 and Mm IPT5-6 p.(Gly1314Val) showing a reduced thermal stability (TWO replicates, see Methods section). Traces correspond to the normalized first derivate of the fluorescence ratio showing the inflection point of the fluorescence ratio, which corresponds to the melting temperature of the sample. T_onset values and protein purification experiments are shown in Supplementary Fig. S5