Extended Data Fig. 5: Raw and analyzed paDSF with OGT, catalytic domains and TPR domain.
From: Protein-adaptive differential scanning fluorimetry using conformationally responsive dyes
(a) Structural model of full-length OGT, based on crystal structures of the TPR (PDB 1W3B) and catalytic (PDB 5C1D) domains. Cartoons of the (b) full-length OGT construct, which includes the full TPR and catalytic domains, (c) catalytic domain construct, which includes the full catalytic domain and TPR repeats 9–13.5, and (d) TPR domain construct, which includes TPR repeats 1-10. (e–h) paDSF results from the 10 hit dyes for OGT, with raw RFU data displayed alongside fitted models for (E) OGT alone, fitted to DSFworld Model 4 (two sigmoids, with initial RFU (F) catalytic domain alone, fitted to DSFworld Model 2 (one sigmoids, with initial RFU (G) TPR domain alone, fitted to DSFworld Model 2 (one sigmoids, with initial RFU (H) overlay of raw data and final fits for all three constructs. (i) Tmas for the three constructs, extracted using the fits displayed in panels A-D. Data are presented as the mean of three technical replicates +/- standard deviation. No significant (ns) difference was found between Tma OGT-1 and Tma cat, nor Tma OGT-2 and Tma TPR. Significance determined using two-tailed t-test, and a significance threshold of p < 0.01.
