Abstract
NMR ligand-affinity screens are vital to drug discovery, are routinely used to screen fragment-based libraries, and used to verify chemical leads from high-throughput assays and virtual screens. NMR ligand-affinity screens are also a highly informative first step towards identifying functional epitopes of unknown proteins, as well as elucidating the biochemical functions of protein–ligand interaction at their binding interfaces. While simple one-dimensional 1H NMR experiments are capable of indicating binding through a change in ligand line shape, they are plagued by broad, ill-defined background signals from protein 1H resonances. We present an uncomplicated method for subtraction of protein background in high-throughput ligand-based affinity screens, and show that its performance is maximized when phase-scatter correction is applied prior to subtraction.
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Acknowledgments
This work was supported, in part, by funds from the National Institutes of Health (Grant number R21AI081154). The research was performed in facilities renovated with support from the National Institutes of Health (Grant number RR015468-01).
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Worley, B., Sisco, N.J. & Powers, R. Statistical removal of background signals from high-throughput 1H NMR line-broadening ligand-affinity screens. J Biomol NMR 63, 53–58 (2015). https://doi.org/10.1007/s10858-015-9962-3
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DOI: https://doi.org/10.1007/s10858-015-9962-3