TY - JOUR T1 - Functional Validation of Virtual Screening for Novel Agents with General Anesthetic Action at Ligand-Gated Ion Channels JF - Molecular Pharmacology JO - Mol Pharmacol DO - 10.1124/mol.113.087692 SP - mol.113.087692 AU - Stephanie A. Heusser AU - Rebecca J. Howard AU - Cecilia M. Borghese AU - Madeline A. Cullins AU - Torben Broemstrup AU - Ui S. Lee AU - Erik Lindahl AU - Jens Carlsson AU - R. Adron Harris Y1 - 2013/01/01 UR - http://molpharm.aspetjournals.org/content/early/2013/08/15/mol.113.087692.abstract N2 - GABAA receptors play a crucial role in the actions of general anesthetics. The recently published crystal structure of the general anesthetic propofol bound to GLIC, a bacterial homolog of GABAA receptors, provided an opportunity to explore structure-based ligand discovery for pentameric ligand-gated ion channels (pLGICs). We used molecular docking of 153,000 commercially available compounds to identify molecules that interact with the propofol binding site in GLIC. In total, twenty-nine compounds were selected for functional testing on recombinant GLIC and 16 of these compounds modulated GLIC function. Active compounds were also tested on recombinant GABAA receptors, and point mutations around the presumed binding pocket were introduced into GLIC and GABAA receptors to test for binding specificity. The potency of active compounds was only weakly correlated with properties such as lipophilicity or molecular weight. One compound was found to mimic the actions of propofol on GLIC and GABAA, and to be sensitive to mutations that reduce the action of propofol in both receptors. Mutant receptors also provided insight about the position of the binding sites and the relevance of the receptor's conformation for anesthetic actions. Overall, the findings support the feasibility of the use of virtual screening to discover allosteric modulators of pLGICs, and suggest that GLIC is a valid model system to identify novel GABAA receptor ligands. ER -