Accurate quantification of protein concentrations and state populations by 2D 1H-N Q-BEST NMR
Abstract
We introduce 2D Q-BEST NMR as a quantitative approach for determining protein concentrations and relative state populations from a set of three 1 H-15 N correlation spectra. The BEST methodology ensures short overall experimental times and highest sensitivity. The method was validated using ubiquitin as a model system, where a precision of approximately 15-20% was achieved at the level of individual cross-peaks. This precision can be further improved by averaging corrected intensities of well-resolved resonances corresponding to the same protein state. The approach was subsequently applied to the quantification of protein concentrations across multiple samples and to the determination of conformational state populations in proteins exhibiting intrinsic or ligand-induced heterogeneity. In both cases, 2D Q-BEST NMR provides accurate and robust quantification of NMR-visible species, demonstrating its utility as a general spectroscopic tool for quantitative biomolecular NMR analysis.
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BibTeX
@article{ouali2026accurate,
title = {{Accurate quantification of protein concentrations and state populations by 2D 1H-N Q-BEST NMR}},
author = {Yacine Ouali and Beate Bersch and Bernhard Brutscher},
journal = {Journal of Biomolecular NMR},
year = {2026},
month = oct,
doi = {10.1007/s10858-026-00498-x},
eprint = {2610.26017},
archivePrefix = {arXiv},
url = {https://hal.science/hal-05801629v1},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Accurate quantification of protein concentrations and state populations by 2D 1H-N Q-BEST NMR
AU - Yacine Ouali
AU - Beate Bersch
AU - Bernhard Brutscher
PY - 2026
DA - 2026/10/14
JO - Journal of Biomolecular NMR
DO - 10.1007/s10858-026-00498-x
UR - https://hal.science/hal-05801629v1
AB - We introduce 2D Q-BEST NMR as a quantitative approach for determining protein concentrations and relative state populations from a set of three 1 H-15 N correlation spectra. The BEST methodology ensures short overall experimental times and highest sensitivity. The method was validated using ubiquitin as a model system, where a precision of approximately 15-20% was achieved at the level of individual cross-peaks. This precision can be further improved by averaging corrected intensities of well-resolved resonances corresponding to the same protein state. The approach was subsequently applied to the quantification of protein concentrations across multiple samples and to the determination of conformational state populations in proteins exhibiting intrinsic or ligand-induced heterogeneity. In both cases, 2D Q-BEST NMR provides accurate and robust quantification of NMR-visible species, demonstrating its utility as a general spectroscopic tool for quantitative biomolecular NMR analysis.
SN - 0925-2738
SN - 1573-5001
ER -