Extended Data Fig. 7: Thermal shifts with L4 from TW408 paDSF.
From: Protein-adaptive differential scanning fluorimetry using conformationally responsive dyes
paDSF results using dye TW408 to monitor titration of L4 with OGT, catalytic ___domain, or TPR ___domain. (a) Schematic of L4 with the four tested conditions: full OGT, catalytic ___domain, TPR ___domain, and a 1:1 mix of catalytic and TPR ___domain. All constructs were tested at 1 µM final; 1:1 mix had 1 µM catalytic ___domain and 1 mM TPR ___domain. (b) Raw RFU for each construct in the presence of 0–10 µM L4. (c) Comparison of resulting Tmas. Left panel: Tma OGT1, Tma cat:TPR 1, and Tma cat; right panel: Tma OGT2, Tma cat:TPR 2, and Tma TPR. Significant thermal upshifts with L4 treatment observed with Tma OGT2, Tma cat:TPR 2, and Tma TPR (Tma OGT2 spearman estimate = 0,95, p = 2.3e-12; Tma cat:TPR 2 spearman estimate = 0.97, p = 4.7e-15; Tma TPR spearman estimate = 0.96, p = 3.6e-14). Significant thermal downshift with L4 observed in Tma OGT1 (data in Fig. 3d, included here for reference, Tma OGT1 spearman estimate = −0.92, p = 6.2e-11). No dose-responsive thermal shift with L4 observed in Tma cat or Tma cat:TPR 1 (Tma cat spearman estimate = −0.27, p = 0.2; Tma cat:TPR 1 spearman estimate = −0.34, p = 0.09). ns = not significant (p > 0.05); *** p < 0.0001. Statistical significance was determined by two-way t-test. Data are presented as the mean of three technical replicates +/- standard deviation.
