Single-Cell Chemical Proteomics with an Activity-Based Probe: Identification of Low-Copy Membrane Proteins on Primary Neurons

doi:10.1002/anie.201402363

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	Single-Cell Chemical Proteomics with an Activity-Based Probe: Identification of Low-Copy Membrane Proteins on Primary Neurons
	Xu, Fei 1; Zhao, Han 2; Feng, Xiaojun 1; Chen, Linhai3 ; Chen, Dongjuan 1; Zhang, Yang 2; Nan, Fajun3 ; Liu, Jianfeng 2; Liu, Bi-Feng 1
刊名	ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
	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.
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关键词	INDUCED FLUORESCENCE DETECTION ; CODED AFFINITY TAGS ; GABA(B) RECEPTOR ; CAPILLARY-ELECTROPHORESIS ; MASS-SPECTROMETRY ; SIGNALING NETWORKS ; GAMMA ; REVEALS ; TRANSACTIVATION ; CHROMATOGRAPHY
WOS研究方向	Chemistry
语种	英语
出版者	WILEY-V C H VERLAG GMBH
WOS记录号	WOS:000338021600027
内容类型	期刊论文
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.