Single-Cell Chemical Proteomics with an Activity-Based Probe: Identification of Low-Copy Membrane Proteins on Primary Neurons
文献类型：期刊论文
| 作者 | Xu, Fei1; Zhao, Han2; Feng, Xiaojun1; Chen, Linhai3 ; Chen, Dongjuan1; Zhang, Yang2; Nan, Fajun3; Liu, Jianfeng2; Liu, Bi-Feng1
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| 刊名 | ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
 |
| 出版日期 | 2014-06 |
| 卷号 | 53期号:26页码:6730-6733 |
| 关键词 | activity-based probes membrane proteins primary neurons proteomics receptors |
| ISSN号 | 1433-7851 |
| DOI | 10.1002/anie.201402363 |
| 文献子类 | Article |
| 英文摘要 | We propose a novel single-cell chemical proteomics (SCCP) strategy to profile low-abundance membrane proteins in single cells. In this approach, the membrane protein GB1 and its splicing variants were targeted on cultured cell lines and primary neurons using a specifically designed activity-based probe. The functionally labeled single cells were encapsulated in individual buffer droplets on a PDMS microwell array, and were further picked up one at a time and loaded into a capillary electrophoresis system for cell lysis, separation, and laser-induced fluorescence detection of the targeted proteins. The results revealed the expression of GB1 splicing variants in HEK and MEF cells, which was previously only suggested at the transcriptional level. We further applied this method to investigate single primary cells and observed significant heterogeneity among individual mouse cerebellar granule neurons. Interference experiments with GB1 antagonist and agonist validated this observation. |
| WOS关键词 | INDUCED FLUORESCENCE DETECTION ; CODED AFFINITY TAGS ; GABA(B) RECEPTOR ; CAPILLARY-ELECTROPHORESIS ; MASS-SPECTROMETRY ; SIGNALING NETWORKS ; GAMMA ; REVEALS ; TRANSACTIVATION ; CHROMATOGRAPHY |
| 资助项目 | NSFC[21275060] ; NSFC[31130028] ; NSFC[31225011] ; NSFC[91213303] ; National Basic Research Program of China[2011CB910403] ; National Basic Research Program of China[2007CB914203] |
| WOS研究方向 | Chemistry |
| 语种 | 英语 |
| WOS记录号 | WOS:000338021600027 |
| 出版者 | WILEY-V C H VERLAG GMBH |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/277042]  |
| 专题 | 国家新药筛选中心 中科院受体结构与功能重点实验室 新药研究国家重点实验室 |
| 通讯作者 | Nan, Fajun |
| 作者单位 | 1.Huazhong Univ Sci & Technol, Coll Life Sci & Technol, Wuhan Natl Lab Optoelect, Wuhan 430074, Peoples R China; 2.Huazhong Univ Sci & Technol, Coll Life Sci & Technol, Key Lab Mol Biophys MOE, Wuhan 430074, Peoples R China; 3.Chinese Acad Sci, Shanghai Inst Mat Med, State Key Lab Drug Res, Shanghai 201203, Peoples R China |
| 推荐引用方式 GB/T 7714 | Xu, Fei,Zhao, Han,Feng, Xiaojun,et al. Single-Cell Chemical Proteomics with an Activity-Based Probe: Identification of Low-Copy Membrane Proteins on Primary Neurons[J]. ANGEWANDTE CHEMIE-INTERNATIONAL EDITION,2014,53(26):6730-6733. |
| APA | Xu, Fei.,Zhao, Han.,Feng, Xiaojun.,Chen, Linhai.,Chen, Dongjuan.,...&Liu, Bi-Feng.(2014).Single-Cell Chemical Proteomics with an Activity-Based Probe: Identification of Low-Copy Membrane Proteins on Primary Neurons.ANGEWANDTE CHEMIE-INTERNATIONAL EDITION,53(26),6730-6733. |
| MLA | Xu, Fei,et al."Single-Cell Chemical Proteomics with an Activity-Based Probe: Identification of Low-Copy Membrane Proteins on Primary Neurons".ANGEWANDTE CHEMIE-INTERNATIONAL EDITION 53.26(2014):6730-6733. |
入库方式： OAI收割
来源：上海药物研究所
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