Extended Data Fig. 8: OGT oligomerization state is not affected by L4, but L4 treatment increases the temperature sensitivity of OGT catalytic activity.
From: Protein-adaptive differential scanning fluorimetry using conformationally responsive dyes
(a) Histograms of counts of MW from mass photometry, used to calculate molecular weights: OGT: 115 kDa, Catalytic domain; 82 kDa, TPR domain: 43, 86, 129 kDa (monomer, dimer, trimer). (b) OGT catalytic activity with a model substrate peptide (KKKYPGGSTPVSSANMM), with enzymatic activity allowed to proceed at 29, 32, 35, 38, 41, 44, 47, or 50 °C. Activity was measured as RLU using Promega UDP-Glo Assay, with higher RLU values corresponding to higher activity. Comparison of temperature-dependent loss of enzymatic activity (RLU) in the presence of increasing concentrations of L4. Solid lines and dots: OGT-containing samples; Dashed lines and Xs: protein-free control. (c) L4-dependent increase in temperature sensitivity of catalytic activity at 41 °C. In all panels, data are presented as the mean of three technical replicates +/- standard deviation.
