Fig. 6: SAXS and ITC analysis of RhoBAST folding and ligand binding.
From: Structural mechanisms for binding and activation of a contact-quenched fluorophore by RhoBAST
a Dimensionless Kratky plots of RhoBAST WT in the presence of various concentrations of Mg2+ and TMR-DN ligand. b Rg and Dmax of RhoBAST derived from guinier plots and PDDF calculation were plotted as a function of Mg2+ concentration. Each data point represents an independent experiment (n = 1) and the error bars are propagated uncertainties calculated by GNOM. c Thermogram of the ITC experiment of RhoBAST binding to TMR-DN in the presence of 10 mM Mg2+ (top) and overlay of integrated fitted heat plots in various Mg2+ concentrations (bottom); for the arithmetic mean of the KD values and thermodynamic parameters, see Supplementary Table 7. d An overlay of the atomic model for wild-type RhoBAST alone (orange L2) built by FARFAR with the crystal structure of RhoBAST in complex with TMR-DN (gray L2) and U1A. For clarity, the U1A protein is not shown. e–g The theoretical scattering profile (shown as red solid line) calculated from the atomic model of RhoBAST WT was fitted with the experimental scattering profiles (shown as gray dots) of RhoBAST in the absence of Mg2+ (e), in the presence of 0.5 mM (f), or both 10 mM Mg2+ and TMR-DN (g). The error bars are propagated uncertainties calculated by GNOM. Source data for panels a–c and e–g are provided as a Source data file.
