File S1 - Structural Insights into Calcium-Bound S100P and the V Domain of the RAGE Complex

Combined file containing supporting figures and tables. Table S1. Active and passive residues used to define the ambiguous interaction restraints for the docking of S100P with the V domain of RAGE. Table S2. Thermodynamic parameters of the interaction between wild-type or mutant S100P and the V domain of RAGE, as determined by ITC. Kd, dissociation constant; ΔH and ΔS, changes in the enthalpy of binding and entropy of binding, respectively; ΔGbinding, Gibbs free energy of binding; T, temperature in Kelvin; and ΔGbinding = ΔH − TΔS. Figure S1. Intermolecular NOEs between the V domain of RAGE* and S100P. The intermolecular NOE peaks between the V domain of RAGE and S100P were observed in 13C(ω2)-edited, 12C(ω3)-filtered NOESY-HSQC experiments and are represented as strip plots. Figure S2. Scatter plot of the HADDOCK score versus the fraction of native contacts (FCC) for a single cluster generated by HADDOCK. Figure S3. Detailed view of Intermolecular NOEs between residues in the RAGE V domain (green) and S100P (cyan) of the modeled RAGE V domain-S100P complex. Figure S4. Secondary structure characterization of wild-type S100P and S100P mutants. Each protein was measured at a concentration of 32 µM in 20 mM Tris-HCl (pH 7.0), 100 mM NaCl, and 4 mM CaCl2. An average of three far-UV CD spectra scans were recorded for each S100P protein from 195 nm to 260 nm using a JASCO-720 spectropolarimeter. (ZIP)