RTA_JBC_2025
Description
Chemical Shift perturbation analysis of small molecules binding to wild-type RTA.
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The NMR data were acquired on Bruker AVANCE III spectrometers equipped with TCI CryoProbesTM at 16.4T and 298K. To map the binding site of the small molecules (RU-NT-192, 206 and 245), 200 ul of 100 uM C13/N15-labeled RTA was mixed with 10 ul of 10 mM compound dissolved in 100% DMSO. The NMR sample buffer included trace protease inhibitors (Roche protease Inhibitor EDTA free tablets), 50 mM Hepes Buffer, 150 mM NaCl, 1 mM TCEP in 5% D2O/95% H2O at pH 7.5. The amide chemical shifts were measured by acquiring 2D 1H-15N HSQC spectra at 16.4T on the free protein and the inhibitor complex at ~1:5 protein-to-compound ratio.
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Funding
National Institutes of Health
AI178870
National Institutes of Health
GM118302