Specific cell surface labeling of GPCRs using split GFP
文献类型：期刊论文
| 作者 | Jiang, Wen-Xue2; Dong, Xu2; Jiang, Jing2; Yang, Yu-Hong2; Yang, Ju2; Lu, Yun-Bi1; Fang, San-Hua3; Wei, Er-Qing1; Tang, Chun2; Zhang, Wei-Ping1 |
| 刊名 | SCIENTIFIC REPORTS
 |
| 出版日期 | 2016-02-09 |
| 卷号 | 6 |
| 英文摘要 | Specific cell surface labeling is essential for visualizing the internalization processes of G-protein coupled receptors (GPCRs) and for gaining mechanistic insight of GPCR functions. Here we present a rapid, specific, and versatile labeling scheme for GPCRs at living-cell membrane with the use of a split green fluorescent protein (GFP). Demonstrated with two GPCRs, GPR17 and CysLT(2)R, we show that two beta-stands (beta-stands 10 and 11) derived from a superfolder GFP (sfGFP) can be engineered to one of the three extracellular loop of a GPCR. The complementary fragment of sfGFP has nine beta-strands (beta-stands 1-9) that carries the mature fluorophore, and can be proteolytically derived from the full-length sfGFP. Separately the GFP fragments are non-fluorescent, but become fluorescent upon assembly, thus allowing specific labeling of the target proteins. The two GFP fragments rapidly assemble and the resulting complex is extremely tight under non-denaturing conditions, which allows real-time and quantitative assessment of the internalized GPCRs. We envision that this labeling scheme will be of great use for labeling other membrane proteins in various biological and pharmacological applications. |
| WOS标题词 | Science & Technology |
| 类目[WOS] | Multidisciplinary Sciences |
| 研究领域[WOS] | Science & Technology - Other Topics |
| 关键词[WOS] | GREEN FLUORESCENT PROTEIN ; COUPLED RECEPTOR ; LIVING CELLS ; IN-VIVO ; INTERNALIZATION ; TRAFFICKING ; GPR17 |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000369579400001 |
| 公开日期 | 2016-04-26 |
| 源URL | [http://ir.wipm.ac.cn/handle/112942/9171]  |
| 专题 | 武汉物理与数学研究所_磁共振应用研究部 |
| 作者单位 | 1.Zhejiang Univ, Dept Pharmacol, Sch Med, Hangzhou 310058, Zhejiang, Peoples R China 2.Chinese Acad Sci, Wuhan Inst Phys & Math, CAS Key Lab Magnet Resonance Biol Syst, State Key Lab Magnet Resonance & Atom Mol Phys, Wuhan 430071, Hubei Province, Peoples R China 3.Zhejiang Univ, Core Facil, Sch Med, Inst Neurosci, Hangzhou 310058, Zhejiang, Peoples R China |
| 推荐引用方式 GB/T 7714 | Jiang, Wen-Xue,Dong, Xu,Jiang, Jing,et al. Specific cell surface labeling of GPCRs using split GFP[J]. SCIENTIFIC REPORTS,2016,6. |
| APA | Jiang, Wen-Xue.,Dong, Xu.,Jiang, Jing.,Yang, Yu-Hong.,Yang, Ju.,...&Zhang, Wei-Ping.(2016).Specific cell surface labeling of GPCRs using split GFP.SCIENTIFIC REPORTS,6. |
| MLA | Jiang, Wen-Xue,et al."Specific cell surface labeling of GPCRs using split GFP".SCIENTIFIC REPORTS 6(2016). |
入库方式： OAI收割
来源：武汉物理与数学研究所
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