title: Discovery of novel ligands for mouse olfactory receptor MOR42-3 using an in silico screening approach and in vitro validation.
abstract: The ligands for many olfactory receptors remain largely unknown despite successful heterologous expression of these receptors. Understanding the molecular receptive range of olfactory receptors and deciphering the olfactory recognition code are hampered by the huge number of odorants and large number of olfactory receptors, as well as the complexity of their combinatorial coding. Here, we present an in silico screening approach to find additional ligands for a mouse olfactory receptor that allows improved definition of its molecular receptive range. A virtual library of 574 odorants was screened against a mouse olfactory receptor MOR42-3. We selected the top 20 candidate ligands using two different scoring functions. These 40 odorant candidate ligands were then tested in vitro using the Xenopus oocyte heterologous expression system and two-electrode voltage clamp electrophysiology. We experimentally confirmed 22 of these ligands. The candidate ligands were screened for both agonist and antagonist activity. In summary, we validated 19 agonists and 3 antagonists. Two of the newly identified antagonists were of low potency. Several previously known ligands (mono- and dicarboxylic acids) are also confirmed in this study. However, some of the newly identified ligands were structurally dissimilar compounds with various functional groups belonging to aldehydes, phenyls, alkenes, esters and ethers. The high positive predictive value of our in silico approach is promising. We believe that this approach can be used for initial deorphanization of olfactory receptors as well as for future comprehensive studies of molecular receptive range of olfactory receptors.
authors.affiliation: Department of Molecular and Cellular Pharmacology, Miller School of Medicine, University of Miami, Miami, Florida, United States of America.
authors.forename: Selvan
authors.initials: S
authors.lastname: Bavan
authors.affiliation: Department of Molecular and Cellular Pharmacology, Miller School of Medicine, University of Miami, Miami, Florida, United States of America.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Sherman
authors.affiliation: Department of Molecular and Cellular Pharmacology, Miller School of Medicine, University of Miami, Miami, Florida, United States of America.
authors.forename: Charles W
authors.initials: CW
authors.lastname: Luetje
authors.affiliation: Department of Molecular and Cellular Pharmacology, Miller School of Medicine, University of Miami, Miami, Florida, United States of America.
authors.forename: Tatjana
authors.initials: T
authors.lastname: Abaffy
mesh_terms: D000818:Animals; D001665:Binding Sites; D016000:Cluster Analysis; D003198:Computer Simulation; D004353:Drug Evaluation, Preclinical; D005260:Female; D008024:Ligands; D051379:Mice; D008958:Models, Molecular; D018035:Receptors, Odorant / Q000819:agonists / Q000378:metabolism*; D015203:Reproducibility of Results; D054852:Small Molecule Libraries / Q000032:analysis* / Q000494:pharmacology*; D014982:Xenopus laevis
chemical_list: D008024:Ligands; D018035:Receptors, Odorant; D054852:Small Molecule Libraries; C000592905:Or55b4 protein, mouse