Time efficient detection of protein–ligand interactions with the polarization optimized PO-WaterLOGSY NMR experiment
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The identification of compounds that bind to a protein of interest is of central importance in contemporary drug research. For screening of compound libraries, NMR techniques are widely used, in particular the Water-Ligand Observed via Gradient SpectroscopY (WaterLOGSY) experiment. Here we present an optimized experiment, the polarization optimized WaterLOGSY (PO-WaterLOGSY). Based on a water flip-back strategy in conjunction with model calculations and numerical simulations, the PO-WaterLOGSY is optimized for water polarization recovery. Compared to a standard setup with the conventional WaterLOGSY, time consuming relaxation delays have been considerably shortened and can even be omitted through this approach. Furthermore, the robustness of the pulse sequence in an industrial setup was increased by the use of hard pulse trains for selective water excitation and water suppression. The PO-WaterLOGSY thus yields increased time efficiency by factor of 3–5 when compared with previously published schemes. These time savings have a substantial impact in drug discovery, since significantly larger compound libraries can be tested in screening campaigns.
KeywordsDrug design NMR screening Protein–ligand interactions WaterLOGSY Water handling
We would like to thank Sebiastian Hiller, Claudio Dalvit and Helena Kovacs for stimulating discussions, and Sandra Jacob and Hans Widmer for critical reading of the manuscript. The authors are indebted to Alain Dietrich for his excellent technical support.
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