Detection of Human GPCR Activity in Drosophila S2 Cells Using the Tango System
G protein-coupled receptors (GPCRs) are essential cell surface proteins involved in transducing extracellular signals into intracellular responses, regulating various physiological processes. This study validated the use of the Tango assay, a sensitive method for detecting GPCR activation, in <i>Drosophila</i> Schneider 2 (S2) cells, focusing on the human Dopamine Receptor D4 (DRD4). Plasmids encoding the LexA-tagged human DRD4 receptor and a luciferase reporter were co-transfected into <i>Drosophila</i> S2 cells and stimulated with dopamine. Receptor activation was measured by quantifying the luciferase activity. The system showed high specificity for dopamine, with no activation in response to octopamine, a non-ligand for DRD4. Furthermore, the system effectively detects activation by a novel compound. These results demonstrate that <i>Drosophila</i> S2 cells, coupled with the Tango assay, provide a viable model for studying human GPCR function and ligand specificity. This system enables the rapid screening of potential GPCR ligands in a cost-effective cellular model.
Authors
- Marianne Marianne
- Andre Rizky Pratomo (ORCID: https://orcid.org/0009-0004-1998-6233)
- Emil Salim (ORCID: https://orcid.org/0000-0002-4667-6351)
- Takayuki Kuraishi (ORCID: https://orcid.org/0000-0002-9493-6082)
- Armia Syahputra (ORCID: https://orcid.org/0000-0002-6474-897X)
- Dadang Irfan Husori (ORCID: https://orcid.org/0000-0001-9003-0526)
- Maryam Aisyah Abdullah (ORCID: https://orcid.org/0009-0009-7238-2160)
- Kamal Rullah (ORCID: https://orcid.org/0000-0001-6856-9373)
- Kohei Matsubara (ORCID: https://orcid.org/0000-0002-2734-7595)
- Nur Farisya Shamsudin (ORCID: https://orcid.org/0000-0002-1167-6618)
- Toshiyuki Takano‐Shimizu (ORCID: https://orcid.org/0000-0002-7058-1256)
- Asuka Inoue (ORCID: https://orcid.org/0000-0003-0805-4049)
- Aki Hori (ORCID: https://orcid.org/0000-0001-5375-6678)
Institutions
- Kanazawa University (JP)
- Tohoku University (JP)
- Kyoto University (JP)
- Universitas Sumatera Utara (ID)
- Kyoto Institute of Technology (JP)
- International Islamic University Malaysia (MY)
Publication Details
- Journal
- International Journal of Molecular Sciences
- Published
- 2024-12-29
- DOI
- https://doi.org/10.3390/ijms26010202
- Primary Topic
- Receptor Mechanisms and Signaling
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Universitas Sumatera Utara
- Japan Science and Technology Agency