Decoding Atomic Addresses: Solution NMR Resonance Assignment of Proteins

Abstract

NMR is a powerful analytical technique that permits the exploration of biomolecules under physiological conditions with atomic resolution. It is especially applicable for examining protein structures and their interactions and dynamics in environments closely resembling their native state, extending its utility to uniquely study disordered proteins. Nevertheless, to extract atomic resolution details, one must successfully correlate observed resonances with their originating nuclei, a process known as ‘resonance assignment’. Even with over fifty years of technical advancements, resonance assignment frequently becomes a bottleneck in the utilization of NMR for the comprehensive study of structure, dynamics, and interactions. In this context, we delve into both the traditional methods and the emerging frontiers in protein resonance assignment strategies for solution NMR. Our goal is to provide a comprehensive view of the existing experimental methodologies, with a focused discussion on their strengths and potential limitations. In this chapter, we will strictly focus on resonance assignment strategies for proteins.