Research into the biological role of the Ca2+-releasing second messenger NAADP (nicotinic acid adenine dinucleotide phosphate) has been hampered by a lack of chemical probes. To find new chemical probes for exploring NAADP signaling, we turned to virtual screening, which can evaluate millions of molecules rapidly and inexpensively. We used NAADP as the query ligand to screen the chemical library ZINC for compounds with similar three-dimensional shape and electrostatic properties. We tested the top-ranking hits in a sea urchin egg bioassay and found that one hit, Ned-19, blocks NAADP signaling at nanomolar concentrations. In intact cells, Ned-19 blocked NAADP signaling and fluorescently labeled NAADP receptors. Moreover, we show the utility of Ned-19 as a chemical probe by using it to demonstrate that NAADP is a key causal link between glucose sensing and Ca2+ increases in mouse pancreatic beta cells.
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Our research was supported by a grant from the Biotechnology and Biological Sciences Research Council (grant number BB/D012694/1). We thank P. Hawkins (OpenEye Scientific Software) for advice with virtual screening, H.-C. Lee (University of Hong Kong) for providing ADP-ribosyl cyclase and C. Garnham (Oxford University) for help with the plate reader.
The authors are going to attempt to patent the diastereomers of Ned-19.
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Naylor, E., Arredouani, A., Vasudevan, S. et al. Identification of a chemical probe for NAADP by virtual screening. Nat Chem Biol 5, 220–226 (2009). https://doi.org/10.1038/nchembio.150
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