Method of making an aromagraph comprising ectopic olfactory receptors
Biosensors are disclosed for detecting ligand binding at ectopic Olfactory Receptors. Methods of identifying novel ectopic Olfactory Receptors are also disclosed. Ligands for ectopic Olfactory Receptors are disclosed as well as methods for using these ligands to interact with ectopic Olfactory Receptors, including the use of such ligands in the treatment and/or mitigation of disease conditions.
1 . A method for making an Aromagraph, comprising the steps of:
providing a plurality of biosensors wherein each biosensor comprises an ectopic Olfactory Receptor, a G protein subunit Ga, a G protein subunit Gβ, and a G protein subunit Gγ, and a reporter, wherein binding of the ligand to the ectopic Olfactory Receptor sends a signal through the G protein subunits resulting in a signal from the reporter, wherein at least some of the biosensors have a different ectopic Olfactory Receptor;
adding the ligand to the biosensors;
detecting an increase for a reporter whereby binding of the ligand to the ectopic Olfactory Receptor is detected, and recording the reporter activity for the plurality of ectopic Olfactory Receptors; and
recording the reporter activity for the plurality of ectopic Olfactory Receptors,
wherein the ectopic Olfactory is expressed on a human kidney cell; and
the biosensor is in a membrane fraction of a host cell,
wherein the plurality of biosensors comprise a plurality of different ectopic Olfactory Receptors.
2 . The method of claim 1 , further comprising the step of quantifying the binding of the ligand to the ectopic Olfactory Receptor.
3 . The method of claim 1 , further comprising the step of quantifying the binding of the ligand at each of the different ectopic Olfactory Receptors.
4 . The method of claim 1 , wherein the reporter is an optical reporter.
5 . The method of claim 4 , further comprising the step of quantifying the binding of the ligand at each of the different ectopic Olfactory Receptors in real time by measuring changes in the optical reporter over time.
6 . The method of claim 1 , wherein the ectopic Olfactory Receptor is selected from the group consisting of an OR51E2, an OR2W3, an OR4N4, an OR51E1, an OR52N4, an OR13A1, an OR5K2, an OR7D2, an OR3A2, an OR2V1, an OR2H2, an OR7C1, an OR2L13, an OR1L8, an OR2T8, an OR10D1, an OR3A3, an OR2K2, an OR13J1, an OR2C1, an OR7A5, an OR10A2, and an OR1F12.