## B_sym1

### Two-site, symmetry test Equilibrium molar fractions of species Other graphic formats: EPS, MATLAB Figure Points correspond to steps in titration series corresponding to Rtotal and LRratio used in NMR line shape simulations below. Curves are smooth functions simulated at constant Rtotal. Populations of multimeric species are shown per monomer. Population of free ligand is normalized to total receptor concentration. Isothermal Titration Calorimetry profile Other graphic formats: EPS, MATLAB Figure Heat uptake curve is simulated as a derivative of the population of species multiplied by specific dH. B_sym1Two-site, symmetry testEquilibrium constants:Ka(A1a)=1.00e+05Ka(A1b)=2.00e+04Ka(A2a)=2.00e+04Ka(A2b)=1.00e+05 (dependent)Rate constants (1-forward, 2-reverse):k1(A1a)=1.00e+05k2(A1a)=1.00e+00k1(A1b)=2.00e+04k2(A1b)=1.00e+00k1(A2a)=2.00e+04k2(A2a)=1.00e+00k1(A2b)=1.00e+05k2(A2b)=1.00e+00Chemical shifts:w0(R)=0.0 /s (0.0 Hz)w0(LaR)=100.0 /s (15.9 Hz)w0(RLb)=200.0 /s (31.8 Hz)w0(RL2)=300.0 /s (47.8 Hz)Base relaxation rates:R2(R)=10.0 /sR2(LaR)=10.0 /sR2(RLb)=10.0 /sR2(RL2)=10.0 /sEnthalpy difference from the base state:dH(R)=0.0dH(LaR)=-2.0dH(RLb)=-1.0dH(RL2)=-3.0Total R concentration (*1000):1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00Ratio of total L to total R:0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 1D-spectra of the titration series Other graphic formats: EPS, MATLAB Figure Traces (bottom to top) correspond to L/R ratios indicated by points in the Populations graph (above). Red dots indicate peak maxima for a visual aid (also plotted as the chemical shift titration curves on the right). Dashed lines indicate chemical shifts of individual species. Chemical shift titration curves Other graphic formats: EPS, MATLAB Figure Points correspond to positions of peak maxima numerically detected in the spectral traces. Sometimes not all peaks will be detected and signal/noise here is not considered.

## B_sym2

### Two-site, symmetry test Equilibrium molar fractions of species Other graphic formats: EPS, MATLAB Figure Points correspond to steps in titration series corresponding to Rtotal and LRratio used in NMR line shape simulations below. Curves are smooth functions simulated at constant Rtotal. Populations of multimeric species are shown per monomer. Population of free ligand is normalized to total receptor concentration. Isothermal Titration Calorimetry profile Other graphic formats: EPS, MATLAB Figure Heat uptake curve is simulated as a derivative of the population of species multiplied by specific dH. B_sym2Two-site, symmetry testEquilibrium constants:Ka(A1a)=2.00e+04Ka(A1b)=1.00e+05Ka(A2a)=1.00e+05Ka(A2b)=2.00e+04 (dependent)Rate constants (1-forward, 2-reverse):k1(A1a)=2.00e+04k2(A1a)=1.00e+00k1(A1b)=1.00e+05k2(A1b)=1.00e+00k1(A2a)=1.00e+05k2(A2a)=1.00e+00k1(A2b)=2.00e+04k2(A2b)=1.00e+00Chemical shifts:w0(R)=0.0 /s (0.0 Hz)w0(LaR)=100.0 /s (15.9 Hz)w0(RLb)=200.0 /s (31.8 Hz)w0(RL2)=300.0 /s (47.8 Hz)Base relaxation rates:R2(R)=10.0 /sR2(LaR)=10.0 /sR2(RLb)=10.0 /sR2(RL2)=10.0 /sEnthalpy difference from the base state:dH(R)=0.0dH(LaR)=-1.0dH(RLb)=-2.0dH(RL2)=-3.0Total R concentration (*1000):1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00Ratio of total L to total R:0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 1D-spectra of the titration series Other graphic formats: EPS, MATLAB Figure Traces (bottom to top) correspond to L/R ratios indicated by points in the Populations graph (above). Red dots indicate peak maxima for a visual aid (also plotted as the chemical shift titration curves on the right). Dashed lines indicate chemical shifts of individual species. Chemical shift titration curves Other graphic formats: EPS, MATLAB Figure Points correspond to positions of peak maxima numerically detected in the spectral traces. Sometimes not all peaks will be detected and signal/noise here is not considered.

LineShapeKin Simulation
Evgenii Kovrigin, Medical College of Wisconsin, 2009