Development of a universal high-throughput calcium assay for G-protein-coupled receptors with promiscuous G-protein G alpha 15/16
文献类型：期刊论文
| 作者 | Zhu, Ting1,2; Fang, Li-yan2,3; Xie, Xin2,3
|
| 刊名 | ACTA PHARMACOLOGICA SINICA
 |
| 出版日期 | 2008-04 |
| 卷号 | 29期号:4页码:507-516 |
| 关键词 | G-protein-coupled receptors G-protein G alpha 15/16 high-throughput screening calcium assay GTP gamma S binding |
| ISSN号 | 1671-4083 |
| DOI | 10.1111/j.1745-7254.2008.00775.x |
| 文献子类 | Article |
| 英文摘要 | Aim: To develop a universal high-throughput screening assay based on G alpha 15/16-mediated calcium mobilization for the identification of novel modulators of G-protein-coupled receptors (GPCR). Methods: In the present study, CHO-K1 or HEK293 cells were co-transfected with plasmids encoding promiscuous G-protein Ga15/16 and various receptors originally coupled to G alpha s, G alpha i, or G alpha q pathways. Intracellular calcium change was monitored with fluorescent dye Fluo-4. Results: We found out for all the receptors tested, G alpha 15/16 could shift the receptors'coupling to the calcium mobilization pathway, and the EC50 values of the ligands generated with this method were comparable with reported values that were obtained using traditional methods. This assay was validated and optimized with the delta-opioid receptor, which originally coupled to G alpha i and was recently found to play important roles in neurodegenerative and autoimmune diseases. A large-scale screening of 48 000 compounds was performed based on this system. Several new modulators were identified and confirmed with the traditional GTP gamma S binding assay. Conclusion: This cell-based calcium assay was proved to be robust and easy to automate, and could be used as a universal method in searching for GPCR modulators. |
| WOS关键词 | DELTA-OPIOID RECEPTOR ; MOLECULAR PHARMACOLOGY ; INTRACELLULAR CALCIUM ; CHEMOKINE RECEPTOR ; DRUG DISCOVERY ; ACTIVATION ; PHOSPHORYLATION ; DESENSITIZATION ; TRANSDUCTION ; MOBILIZATION |
| WOS研究方向 | Chemistry ; Pharmacology & Pharmacy |
| 语种 | 英语 |
| WOS记录号 | WOS:000254559000016 |
| 出版者 | BLACKWELL PUBLISHING |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/272944]  |
| 专题 | 国家新药筛选中心 |
| 通讯作者 | Xie, Xin |
| 作者单位 | 1.Chinese Acad Sci, Shanghai Inst Biol Sci, Shanghai 201203, Peoples R China 2.Chinese Acad Sci, Natl Ctr Drug Screening, Shanghai 201203, Peoples R China; 3.Chinese Acad Sci, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China; |
| 推荐引用方式 GB/T 7714 | Zhu, Ting,Fang, Li-yan,Xie, Xin. Development of a universal high-throughput calcium assay for G-protein-coupled receptors with promiscuous G-protein G alpha 15/16[J]. ACTA PHARMACOLOGICA SINICA,2008,29(4):507-516. |
| APA | Zhu, Ting,Fang, Li-yan,&Xie, Xin.(2008).Development of a universal high-throughput calcium assay for G-protein-coupled receptors with promiscuous G-protein G alpha 15/16.ACTA PHARMACOLOGICA SINICA,29(4),507-516. |
| MLA | Zhu, Ting,et al."Development of a universal high-throughput calcium assay for G-protein-coupled receptors with promiscuous G-protein G alpha 15/16".ACTA PHARMACOLOGICA SINICA 29.4(2008):507-516. |
入库方式： OAI收割
来源：上海药物研究所
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