We have further developed our global multi-method analysis approach, which combines independent observation of different observables from different techniques. We have further improved our data analysis program SEDPHAT for this purpose. It currently allows the seamless combination of sedimentation velocity, sedimentation equilibrium, dynamic light scattering, isothermal titration calorimetry, surface binding, and different spectroscopies. Key new developments are the statistical tools of global analysis. We have developed consistency matrices, which report to what degree experimental data sets are consistent and/or contribute new orthogonal information, and information content matrices, which report the improvement in binding parameters arising from the inclusion of each data set. Furthermore, we improved our strategies for weighting different data sets. We have applied these tools to a comprehensive model system with two-site binding, studied by surface plasmon resonance, isothermal titration calorimetry, sedimentation velocity, and fluorescence polarization. Since isothermal titration calorimetry is a uniquely useful tool for the study of macromolecular binding energetics, we have embarked on the improvement of data quality in this technique. In collaboration with Dr. Chad Brautigam, we have developed a noval approach for extracting heats of reaction at each titration step from the time course of the power signal that constitutes the raw recorded data, based on peak-shape analysis and singular value decomposition. This resulted in a new public domain software NITPIC, which was developed as a standalone tool for the integration of thermograms.
