Heteronuclear adiabatic relaxation dispersion (hard) for quantitative analysis of conformational dynamics in proteins

Nathaniel Traaseth, Fa An Chao, Larry R. Masterson, Silvia Mangia, Michael Garwood, Shalom Michaeli, Burckhard Seelig, Gianluigi Veglia

Research output: Contribution to journalArticle

Abstract

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 R 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 R and R 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 R and R 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.

Original languageEnglish (US)
Pages (from-to)75-82
Number of pages8
JournalJournal of Magnetic Resonance
Volume219
DOIs
StatePublished - Jun 2012

Fingerprint

quantitative analysis
proteins
Chemical analysis
Proteins
Rate constants
Nuclear magnetic resonance
Biomolecules
Ligases
Ubiquitin
Catalysis
Radio
nuclear magnetic resonance
probes
Experiments
complement
RNA
folding
catalysis
dynamic range
enzymes

Keywords

  • Adiabatic relaxation dispersion
  • NMR
  • Proteins
  • Rotating frame relaxation

ASJC Scopus subject areas

  • Nuclear and High Energy Physics
  • Biochemistry
  • Biophysics
  • Condensed Matter Physics

Cite this

Heteronuclear adiabatic relaxation dispersion (hard) for quantitative analysis of conformational dynamics in proteins. / Traaseth, Nathaniel; Chao, Fa An; Masterson, Larry R.; Mangia, Silvia; Garwood, Michael; Michaeli, Shalom; Seelig, Burckhard; Veglia, Gianluigi.

In: Journal of Magnetic Resonance, Vol. 219, 06.2012, p. 75-82.

Research output: Contribution to journalArticle

Traaseth, Nathaniel ; Chao, Fa An ; Masterson, Larry R. ; Mangia, Silvia ; Garwood, Michael ; Michaeli, Shalom ; Seelig, Burckhard ; Veglia, Gianluigi. / Heteronuclear adiabatic relaxation dispersion (hard) for quantitative analysis of conformational dynamics in proteins. In: Journal of Magnetic Resonance. 2012 ; Vol. 219. pp. 75-82.
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AU - Garwood, Michael

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