Multi-Method Approaches for the Study of Complex Protein Interactions

Biomedical Imaging & Bioengineering

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Abstract

Since a single biophysical technique is limited in the number of observable properties, one promising approach is the simultaneous consideration of data from multiple biophysical techniques. Therefore, one important goal is development of a robust and general computational framework, conducted with different techniques, for the global analysis of binding studies of triple (or higher) protein mixtures. In the past, we have developed for this purpose a data analysis program SEDPHAT for the global analysis of data from sedimentation velocity, sedimentation equilibrium, dynamic light scattering, isothermal titration calorimetry, and surface binding. It is now widely used in the biophysical community.[unreadable] [unreadable] Guided by experimental systems studied collaboratively with LCMB/NCI, we have implemented several binding models for multiple cooperative triple protein interactions. We have further explored the capability of isothermal titration calorimetry as an experimental method to study cooperativity, introduced novel global modelling approaches, and devised new experimental calorimetry protocols. We have identified several protein interactions that can serve as model systems for the study of the detailed compatibility of data from surface binding, calorimetry, circular dichroism spectroscopy, and sedimentation, and have acquired experimental data. This will be crucial to establishing the level of detail on the macromolecular interactions that a global analysis is able to extract.