Characterizing micro-to-millisecond chemical exchange in nucleic acids using off-resonance R1ρ relaxation dispersion

This review describes off-resonance R-1 rho relaxation dispersion NMR methods for characterizing microsecond-to-millisecond chemical exchange in uniformly C-13/N-15 labeled nucleic acids in solution. The review opens with a historical account of key developments that formed the basis for modern R-1 rho techniques used to study chemical exchange in biomolecules. A vector model is then used to describe the R-1 rho relaxation dispersion experiment, and how the exchange contribution to relaxation varies with the amplitude and frequency offset of an applied spin-locking field, as well as the population, exchange rate, and differences in chemical shifts of two exchanging species. Mathematical treatment of chemical exchange based on the Bloch-McConnell equations is then presented and used to examine relaxation dispersion profiles for more complex exchange scenarios including three-state exchange. Pulse sequences that employ selective Hartmann-Hahn cross-polarization transfers to excite individual C-13 or N-15 spins are then described for measuring off-resonance R-1 rho(C-13) and R-1 rho(N-15) in uniformly C-13/N-15 labeled DNA and RNA samples prepared using commercially available C-13/N-15 labeled nucleotide triphosphates. Approaches for analyzing R-1 rho data measured at a single static magnetic field to extract a full set of exchange parameters are then presented that rely on numerical integration of the Bloch-McConnell equations or the use of algebraic expressions. Methods for determining structures of nucleic acid excited states are then reviewed that rely on mutations and chemical modifications to bias conformational equilibria, as well as structure-based approaches to calculate chemical shifts. Applications of the methodology to the study of DNA and RNA conformational dynamics are reviewed and the biological significance of the exchange processes is briefly discussed. (C) 2019 Elsevier B.V. All rights reserved.