Boosting the resolution of low-field $$^{15}\hbox {N}$$15N relaxation experiments on intrinsically disordered proteins with triple-resonance NMR

[nmrlearner]

Boosting the resolution of low-field $$^{15}\hbox {N}$$15N relaxation experiments on intrinsically disordered proteins with triple-resonance NMR

Abstract

Improving our understanding of nanosecond motions in disordered proteins requires the enhanced sampling of the spectral density function obtained from relaxation at low magnetic fields. High-resolution relaxometry and two-field NMR measurements of relaxation have, so far, only been based on the recording of one- or two-dimensional spectra, which provide insufficient resolution for challenging disordered proteins. Here, we introduce a 3D-HNCO-based two-field NMR experiment for measurements of protein backbone \(^{15}\hbox {N}\) amide longitudinal relaxation rates. The experiment provides accurate longitudinal relaxation rates at low field (0.33 T in our case) preserving the resolution and sensitivity typical for high-field NMR spectroscopy. Radiofrequency pulses applied on six different radiofrequency channels are used to manipulate the spin system at both fields. The experiment was demonstrated on the C-terminal domain of \(\delta\) subunit of RNA polymerase from Bacillus subtilis, a protein with highly repetitive amino-acid sequence and very low dispersion of backbone chemical shifts.



Source: Journal of Biomolecular NMR