Abstract
Odorant responsiveness of a mouse olfactory receptor, mOR-EG, was investigated in various heterologous cells using a variety of detection methods. Odorant-induced Ca2+ response was observed in HEK293 cells that coexpressed mOR-EG and the promiscuous G protein, Gα15. Without Gα15, a robust increase in cAMP level was observed upon odorant-stimulation in various mammalian cells. A luciferase reporter gene assay using zif268 promoter was adopted to amplify the cAMP signals. In Xenopus laevis oocytes, odorant-stimulated currents were recorded when mOR-EG cRNA was co-injected with either Gα15 or cAMP-dependent channel. These results suggest that odorant responsiveness can be monitored via a signaling pathway mediated by endogenous Gαs or transfected Gα15 in heterologous cell systems. Various functional assays for a heterologously expressed olfactory receptor reported in this study, are potentially useful for high-throughput ligand screening and functional analyses of hundreds of olfactory receptors.
| Original language | English |
|---|---|
| Pages (from-to) | 964-969 |
| Number of pages | 6 |
| Journal | Biochemical and Biophysical Research Communications |
| Volume | 305 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - Jun 13 2003 |
Fingerprint
All Science Journal Classification (ASJC) codes
- Biophysics
- Biochemistry
- Molecular Biology
- Cell Biology
Cite this
Odorant response assays for a heterologously expressed olfactory receptor. / Katada, Sayako; Nakagawa, Takao; Kataoka, Hiroshi; Touhara, Kazushige.
In: Biochemical and Biophysical Research Communications, Vol. 305, No. 4, 13.06.2003, p. 964-969.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Odorant response assays for a heterologously expressed olfactory receptor
AU - Katada, Sayako
AU - Nakagawa, Takao
AU - Kataoka, Hiroshi
AU - Touhara, Kazushige
PY - 2003/6/13
Y1 - 2003/6/13
N2 - Odorant responsiveness of a mouse olfactory receptor, mOR-EG, was investigated in various heterologous cells using a variety of detection methods. Odorant-induced Ca2+ response was observed in HEK293 cells that coexpressed mOR-EG and the promiscuous G protein, Gα15. Without Gα15, a robust increase in cAMP level was observed upon odorant-stimulation in various mammalian cells. A luciferase reporter gene assay using zif268 promoter was adopted to amplify the cAMP signals. In Xenopus laevis oocytes, odorant-stimulated currents were recorded when mOR-EG cRNA was co-injected with either Gα15 or cAMP-dependent channel. These results suggest that odorant responsiveness can be monitored via a signaling pathway mediated by endogenous Gαs or transfected Gα15 in heterologous cell systems. Various functional assays for a heterologously expressed olfactory receptor reported in this study, are potentially useful for high-throughput ligand screening and functional analyses of hundreds of olfactory receptors.
AB - Odorant responsiveness of a mouse olfactory receptor, mOR-EG, was investigated in various heterologous cells using a variety of detection methods. Odorant-induced Ca2+ response was observed in HEK293 cells that coexpressed mOR-EG and the promiscuous G protein, Gα15. Without Gα15, a robust increase in cAMP level was observed upon odorant-stimulation in various mammalian cells. A luciferase reporter gene assay using zif268 promoter was adopted to amplify the cAMP signals. In Xenopus laevis oocytes, odorant-stimulated currents were recorded when mOR-EG cRNA was co-injected with either Gα15 or cAMP-dependent channel. These results suggest that odorant responsiveness can be monitored via a signaling pathway mediated by endogenous Gαs or transfected Gα15 in heterologous cell systems. Various functional assays for a heterologously expressed olfactory receptor reported in this study, are potentially useful for high-throughput ligand screening and functional analyses of hundreds of olfactory receptors.
UR - http://www.scopus.com/inward/record.url?scp=0038242292&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0038242292&partnerID=8YFLogxK
U2 - 10.1016/S0006-291X(03)00863-5
DO - 10.1016/S0006-291X(03)00863-5
M3 - Article
C2 - 12767924
AN - SCOPUS:0038242292
VL - 305
SP - 964
EP - 969
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 4
ER -