Sedimentation Velocity with Fluorescence Detection (SV-FDS) and its Applications to the Studies of Self- and Hetero-Associations of Ionotropic Glutamate Receptor Domains

Authors

• H ZHAO
• S Lomash
• C Glasser
• S Chittori
• M Mayer
• P Schuck

Abstract

Sedimentation velocity (SV) analytical ultracentrifugation (AUC) has been widely applied to study macromolecules in free solution to resolve their size and shape, as well as their reversible interactions. With the new fluorescence detection system (FDS) being recently commercially available, SV-AUC has become particularly powerful for quantitatively measure the binding of molecules with high affinity, such as Kd in nM and even pM regime. Here we demonstrate the exclusive advantages offered by FDS-SV for investigating protein-protein interactions and its application to quantify a series of specific self and hetero-association between amino terminal domains of different ionotropic glutamate receptor (iGluR ATD). FDS-SV not only reveals the preferential binding nature for certain pairs, but also provides kinetic information on the interactions.

Scientific Focus Area: Biomedical Engineering and Biophysics

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