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Ligand‐Detected Relaxation Dispersion NMR Spectroscopy: Dynamics of preQ 1 –RNA Binding
Author(s) -
Moschen Thomas,
Wunderlich Christoph Hermann,
Spitzer Romana,
Levic Jasmin,
Micura Ronald,
Tollinger Martin,
Kreutz Christoph
Publication year - 2015
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201409779
Subject(s) - riboswitch , chemistry , ligand (biochemistry) , receptor–ligand kinetics , biomolecule , relaxation (psychology) , nuclear magnetic resonance spectroscopy , population , rna , dispersion (optics) , small molecule , aptamer , molecule , kinetics , biophysics , chemical physics , stereochemistry , receptor , non coding rna , biochemistry , biology , physics , genetics , organic chemistry , demography , quantum mechanics , neuroscience , sociology , optics , gene
An NMR‐based approach to characterizing the binding kinetics of ligand molecules to biomolecules, like RNA or proteins, by ligand‐detected Carr‐Purcell‐Meiboom‐Gill (CPMG) relaxation dispersion experiments is described. A 15 N‐modified preQ 1 ligand is used to acquire relaxation dispersion experiments in the presence of low amounts of the Fsu class I preQ 1 aptamer RNA, and increasing ligand concentrations to probe the RNA small molecule interaction. Our experimental data strongly support the conformational selection mechanism postulated. The approach gives direct access to two parameters of a ligand–receptor interaction: the off rate and the population of the small molecule–receptor complex. A detailed description of the kinetics underlying the ligand binding process is of crucial importance to fully understanding a riboswitch’s function and to evaluate potential new antibiotics candidates targeting the noncoding RNA species. Ligand‐detected NMR relaxation dispersion experiments represent a valuable diagnostic tool for the characterization of binding mechanisms.