Experimental techniques employed are analytical ultracentrifugation, static and dynamic light scattering, isothermal titration microcalorimetry, differential scanning calorimetry, circular dichroism spectroscopy, fluorescence polarization, and surface plasmon resonance biosensing. Specifically, in collaboration with Dr. Mark Mayer, we have applied sedimentation velocity to the high-affinity interactions of glutamate receptor subunits, in particular GluA2. Detailed methodological comparisons were made between sedimentation velocity, sedimentation equilibrium and steady-state fluorescence anisotropy, and fluorescence detected sedimentation velocity when applied to high-affinity interactions. This will be critical for future characterization of homo- and heterodimerization of glutamate receptor subunits. We have implemented new sedimentation velocity methodology to improve the study of the high-affinity interaction of Swi6 oligomerization and histone binding, jointly with Dr. Geeta Narlikar. This required the development of an isodesmic association model, and its application in the global analysis of sedimentation velocity and sedimentation equilibrium. New sedimentation velocity methods were also implemented to study a trimeric TRAP transporter and its interactions, in collaboration with Dr. Chad Brautigam. Finally, we have continued our collaboration with Dr. Graeme Wistow on the experimental biophysical characterization of hydration and weak interactions of gamma crystallins from different species, and we have collaborated with Dr. Geoffrey Howlett in the developed equilibrium models for the length distribution of amyloid fibrils.
NIH ExPorter
NIH ExPorter