Olfactory receptors for isolvaleric acid and related malodorants and use thereof in assays for identification of blockers

Abstract

A subgenus of olfactory receptors (ORs) that are activated by isovaleric acid (IVA) are identified as well as assays that utilize one or more of these ORs. These assays are useful for identifying potential anti-odorants which may be used in deodorants, air and carpet fresheners, fabric deodorizers, and other compositions for camouflaging odor attributable to IVA and related carboxylic acids.

1 - 58 . (canceled) 
     
     
         59 . A method of screening for a compound that putatively blocks isovaleric acid associated malodor comprising:
 (i) contacting an isovaleric acid olfactory receptor polypeptide that possesses at least 95% sequence identity to the polypeptide contained in SEQ ID NO:24 with at least one compound;   (ii) assaying whether said at least one compound inhibits the binding of said olfactory receptor polypeptide to isovaleric acid; and   (iii) identifying a compound that putatively blocks isovaleric associated malodor if it inhibits the said binding of said olfactory receptor polypeptide to isovaleric acid.   
     
     
         60 . The screening method of  claim 59  wherein said identified compound is assessed in a smell test to determine whether said identified compound blocks or inhibits isovaleric acid associated malodor. 
     
     
         61 . The screening method of  claim 59  wherein the olfactory receptor polypeptide has the sequence contained in SEQ ID NO:24. 
     
     
         62 . The method of  claim 59  wherein said olfactory receptor polypeptide is expressed by a cell that also expresses a G protein. 
     
     
         63 . The method of  claim 62  wherein the G protein is Galpha15 or Galpha16. 
     
     
         64 . The screening method of  claim 62  wherein the cell is selected from a HEK-293, COS, and a CHO cell. 
     
     
         65 . The screening method of  claim 62  wherein the cell is bound to a solid phase. 
     
     
         66 . A method of identifying a compound that putatively modulates isovaleric acid associated malodor comprising:
 (i) contacting a cell line that expresses an isovaleric acid receptor polypeptide that has a sequence that is at least 95% identical to SEQ ID NO:24 with at least one compound;   (ii) screening for compounds that block or inhibit the activity of said olfactory receptor polypeptide; and   (iii) identifying a compound that putatively modulates isovaleric acid associated malodor if it blocks or inhibits the activity of said isovaleric acid receptor polypeptide.   
     
     
         67 . The method of  claim 66  wherein the receptor is expressed by a cell that additionally expresses a G protein. 
     
     
         68 . The method of  claim 66  wherein said cell is selected from a  Xenopus  oocyte, CHO cell, HEK-293 cell and a HeLa cell. 
     
     
         69 . The method of  claim 67  wherein the G protein is Galpha15 or Galpha16. 
     
     
         70 . The method of  claim 66  wherein said receptor polypeptide has the sequence contained in SEQ ID NO:24. 
     
     
         71 . The method of  claim 66  wherein the screening assay selects for compounds that affect receptor internalization. 
     
     
         70 . The method of  claim 66  wherein the screening assay selects for compounds that affect receptor phosphorylation. 
     
     
         71 . The method of  claim 66  wherein the screening assay selects for compounds that affect arrestin translocation. 
     
     
         72 . The method of  claim 66  wherein the assay screens for compounds that affect G protein activation of said receptor. 
     
     
         73 . The method of  claim 66  wherein the screening assay selects for compounds that affect the conformation of said olfactory receptor polypeptide. 
     
     
         74 . The method of  claim 73  wherein the screening assay selects for compounds that alter the susceptibility of said receptor polypeptide to proteolysis. 
     
     
         75 . The method of  claim 73  wherein the screening assay detects any change in conformation using NMR spectroscopy. 
     
     
         76 . The method of  claim 73  wherein the screening assay detects any conformational change by fluorescence spectroscopy. 
     
     
         77 . The method of  claim 66  which screens for compounds that displace binding of a radioactively or fluorescently labeled ligand to said olfactory receptor polypeptide. 
     
     
         78 . The method of  claim 77  which detects the displacement of said labeled compound by fluorescence polarization or FRET assay. 
     
     
         79 . The method of  claim 66  which is a high throughput screening assay. 
     
     
         80 . The method of  claim 66  wherein the receptor is attached to a peptide that facilitates surface expression. 
     
     
         81 . The method of  claim 66  wherein a gene encoding the receptor is operably linked to a reporter gene. 
     
     
         82 . The method of  claim 81  wherein the reporter is luciferase, beta galactosidase, alkaline phosphatase or beta lactamase.