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Abstract
Dresden G protein-coupled receptor (D-GPCR) is one of orphan G protein-coupled receptors (GPCR). Here we report the identification of the ligands and the characterization of D-GPCR. We investigated over 5000 compounds to evoke the response mediated by D-GPCR and identified 3-methyl-valeric acid and 4-methyl-valeric acid as agonists using a cAMP assay. It is of interest that they dramatically enhanced the intracellular cAMP accumulation and the CRE-luciferase activity in CHO-K1 cells and HEK293 cells expressing the chimeric protein of D-GPCR with a rhodopsin-tag at its N-terminus. Our results established new characteristics of D-GPCR as an olfactory receptor. First, agonists of D-GPCR belong to odorants. Second, D-GPCR mRNA is expressed in the olfactory bulb. In addition, D-GPCR was reported to have similar sequences and its genome locus nearby other olfactory receptors. These results suggest D-GPCR is an olfactory receptor.
| Abbreviations: | ||
| GPCR | = | G-protein coupled receptor |
| Rho-tag | = | rhodopsin-tag |
| DMEM | = | Dulbecco's Modified Eagle's medium |
| IBMX | = | 3-isobutyl-1-methyl-xanthine |
| CHO | = | Chinese hamster ovary |
| PCR | = | polymerase chain reaction |
| HEK293 | = | human embryonic kidney 293 |
| GFP | = | green fluorescent protein |
| SDS-PAGE | = | sodium dodecyl sulfate polyacrylamide gel electrophoresis |
| PVDF | = | PolyVinylidine DiFluoride |
| PBS | = | phosphate buffered saline |
| cAMP | = | adenosine 3′,5′–cyclic monophosphate |
| EGF | = | epidermal growth factor |
| EC50 | = | medium effective concentration. |
| Abbreviations: | ||
| GPCR | = | G-protein coupled receptor |
| Rho-tag | = | rhodopsin-tag |
| DMEM | = | Dulbecco's Modified Eagle's medium |
| IBMX | = | 3-isobutyl-1-methyl-xanthine |
| CHO | = | Chinese hamster ovary |
| PCR | = | polymerase chain reaction |
| HEK293 | = | human embryonic kidney 293 |
| GFP | = | green fluorescent protein |
| SDS-PAGE | = | sodium dodecyl sulfate polyacrylamide gel electrophoresis |
| PVDF | = | PolyVinylidine DiFluoride |
| PBS | = | phosphate buffered saline |
| cAMP | = | adenosine 3′,5′–cyclic monophosphate |
| EGF | = | epidermal growth factor |
| EC50 | = | medium effective concentration. |