Methods for characterizing conformational dynamics by solid state NMR

PHYS 187

Ann E. McDermott, aem5@columbia.edu, Department of Chemistry, Columbia University, New York, NY 10027
NMR methods to study the conformational dynamics of microcrystalline enzymes will be discussed, with the examples of ubiquitin and Triosephosphate Isomerase. With extensive or complete resonance assignments in hand, fast-limit order parameters can be measured (on the low microsecond or submicrosecond timescale), as well as intermediate exchange motions (on the high microsecond to millisecond timescale). Fast limit motions of ubiquitin detected in these studies are extensive in comparison with prior biophysical studies on a faster timescale. Methods for cross-validation of this conclusion will be discussed. Studies of Triosephosphate Isomerase illustrate coordination of conformational exchange to ligand binding and release.