Chapter 5 - Protein Dynamics as Studied by Solution NMR Techniques

Abstract

Recent advances in the field of high-resolution nuclear magnetic resonance (NMR) spectroscopy have provided a means by which the structure and dynamics of macromolecules in solution can be characterized at atomic resolution. The focus of this chapter is on recently developed two-dimensional (2D) heteronuclear NMR techniques for measurement of nuclear spin relaxation of proteins in solution, and on the interpretation of these measurements to yield a detailed description of global and internal motions. The application of some of these techniques to staphylococcal nuclease (SNase) in the presence and absence of ligands is included to illustrate the advantages and limitations of such studies, and to demonstrate the types of information that is obtained. Proton detected heteronuclear two-dimensional NMR relaxation measurement techniques provide a powerful tool with which to characterize the internal motions of proteins in solution. Herein, the chapter reviews recently developed pulse sequences, which are optimized for measurement of T1, T2, and NOE values for AX and AX3 spin systems. We have illustrated the use of these techniques by application to specific backbone and side chain sites in SNase. Although in this review, we have focused on a single protein, the backbone and side chain dynamics of several other 15N or 13C labeled proteins and peptides are studied using similar techniques.