Advances in solid-state relaxation methodology for probing site-specific protein dynamics.

Dynamics are intimately linked to protein stability and play a crucial role in important biological processes, such as ligand binding, allosteric regulation, protein folding, signaling, and enzymatic catalysis. Solid-state NMR relaxation measurements allow researchers to determine the amplitudes, time scales, and under favorable conditions, directionality of motions at atomic resolution over the entire range of dynamic processes from picoseconds to milliseconds. Because this method allows researchers to examine both the amplitudes and time scales of motions in this range, they can link different tiers of protein motions in protein energy landscapes. As a result, scientists can better understand the relationships between protein motions and functions. Such studies are possible both with the primary targets of solid-state NMR studies, such as amyloid fibrils, membrane proteins, or other heterogeneous systems, and others that researchers typically study by solution NMR and X-ray crystallography. In addition, solid-state NMR, with the absence of tumbling in solution, eliminates the intrinsic size limitation imposed by slow tumbling of large proteins. Thus, this technique allows researchers to characterize interdomain and intermolecular interactions in large complexes at the atomic scale. In this Account, we discuss recent advances in solid-state relaxation methodology for studying widespread site-specific protein dynamics. We focus on applications involving magic angle spinning, one of the primary methods used in high-resolution solid-state NMR. We give an overview of challenges and solutions for measuring (15)N and (13)C spin-lattice relaxation (R1) to characterize fast picosecond-nanosecond motions, spin-lattice in the rotating frame (R1ρ), and other related relaxation rates for characterization of picosecond-millisecond protein motions. In particular, we discuss the problem of separating incoherent effects caused by random motions from coherent effects arising from incomplete averaging of orientation-dependent NMR interactions. We mention a number of quantitative studies of protein dynamics based on solid-state relaxation measurements. Finally, we discuss the potential use of relaxation measurements for extracting the directionality of motions. Using the (15)N and (13)C R1 and R1ρ measurements, we illustrate the backbone and side-chain dynamics in the protein GB1 and comment on this emerging dynamic picture within the context of data from solution NMR measurements and simulations.