title: Identification of a chemical probe for NAADP by virtual screening.
abstract: Research into the biological role of the Ca(2+)-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 Ca(2+) increases in mouse pancreatic beta cells.
authors.affiliation: Department of Pharmacology, University of Oxford, Oxford, UK.
authors.forename: Edmund
authors.initials: E
authors.lastname: Naylor
authors.forename: Abdelilah
authors.initials: A
authors.lastname: Arredouani
authors.forename: Sridhar R
authors.initials: SR
authors.lastname: Vasudevan
authors.forename: Alexander M
authors.initials: AM
authors.lastname: Lewis
authors.forename: Raman
authors.initials: R
authors.lastname: Parkesh
authors.forename: Akiko
authors.initials: A
authors.lastname: Mizote
authors.forename: Daniel
authors.initials: D
authors.lastname: Rosen
authors.forename: Justyn M
authors.initials: JM
authors.lastname: Thomas
authors.forename: Minoru
authors.initials: M
authors.lastname: Izumi
authors.forename: A
authors.initials: A
authors.lastname: Ganesan
authors.forename: Antony
authors.initials: A
authors.lastname: Galione
authors.forename: Grant C
authors.initials: GC
authors.lastname: Churchill
mesh_terms: D000818:Animals; D002243:Carbolines / Q000737:chemistry / Q000494:pharmacology; D036563:Cyclic ADP-Ribose / Q000494:pharmacology; D015544:Inositol 1,4,5-Trisphosphate / Q000494:pharmacology; D050417:Insulin-Secreting Cells / Q000187:drug effects; D051379:Mice; D008958:Models, Molecular; D015394:Molecular Structure; D009249:NADP / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D010063:Ovum / Q000737:chemistry; D010879:Piperazines / Q000737:chemistry / Q000494:pharmacology; D012617:Sea Urchins; D054852:Small Molecule Libraries
chemical_list: C539827:1-(3-((4-(2-fluorophenyl)piperazin-1-yl)methyl)-4-methoxyphenyl)-2,3,4,9-tetrahydro-1H-pyrido(3,4-b)indole-3-carboxylic acid; D002243:Carbolines; D010879:Piperazines; D054852:Small Molecule Libraries; D036563:Cyclic ADP-Ribose; D009249:NADP; C024376:NAADP; D015544:Inositol 1,4,5-Trisphosphate