Use of Protonless NMR Spectroscopy To Alleviate the Loss of Information Resulting from Exchange-Broadening

We report here the use of protonless NMR spectroscopy to extract structural information under biologically relevant conditions when conventional proton-detection NMR spectroscopy fails due to the loss of labile proton resonances. By direct C-13 detection, correlations between nonlabile nuclei of a given biomolecule can be determined with high resolution, which becomes particularly useful when the system of interests is sensitive to solvent exchange at elevated temperatures, such as intrinsically disordered proteins. Human alpha-synuclein, which is associated with Parkinson's disease, is used as a model system to illustrate the usefulness of protonless NMR spectroscopy in recovering hitherto missing spectral. information.