NMR relaxation methods probe biomolecular motions over a wide range of timescales. In particular, the rotating frame spin-lock R1 ρ and Carr-Purcell-Meiboom-Gill (CPMG) R2 experiments are commonly used to characterize μs to ms dynamics, which play a critical role in enzyme folding and catalysis. In an effort to complement these approaches, we introduced the Heteronuclear Adiabatic Relaxation Dispersion (HARD) method, where dispersion in rotating frame relaxation rate constants (longitudinal R1ρ and transverse R2 ρ) is created by modulating the shape and duration of adiabatic full passage (AFP) pulses. Previously, we showed the ability of the HARD method to detect chemical exchange dynamics in the fast exchange regime (kex ∼ 104-105 s-1). In this article, we show the sensitivity of the HARD method to slower exchange processes by measuring R1ρ and R2ρ relaxation rates for two soluble proteins (ubiquitin and 10C RNA ligase). One advantage of the HARD method is its nominal dependence on the applied radio frequency field, which can be leveraged to modulate the dispersion in the relaxation rate constants. In addition, we also include product operator simulations to define the dynamic range of adiabatic R1ρ and R 2ρ that is valid under all exchange regimes. We conclude from both experimental observations and simulations that this method is complementary to CPMG-based and rotating frame spin-lock R1 ρ experiments to probe conformational exchange dynamics for biomolecules. Finally, this approach is germane to several NMR-active nuclei, where relaxation rates are frequency-offset independent.
Bibliographical noteFunding Information:
This work was supported by National Institutes of Health Grants GM072701 (G.V.), BTRR – P41 RR008079 (CMRR), R01NS061866 (S.M.), NASA Astrobiology Institute NNX09AH70A (B.S.).
- Adiabatic relaxation dispersion
- Rotating frame relaxation