A Cis-Membrane FRET-Based Method for Protein-Specific Imaging of   Cell-Surface Glycans

doi:10.1021/ja410086d

CORC > 北京大学 > 化学与分子工程学院

	A Cis-Membrane FRET-Based Method for Protein-Specific Imaging of Cell-Surface Glycans
	Lin, Wei ; Du, Yifei ; Zhu, Yuntao ; Chen, Xing
刊名	journal of the american chemical society
	2014
关键词	TGF-BETA RECEPTOR LINKED GLYCOSYLATION TRPV5 CHANNEL LIVING CELLS ACTIVATION INTEGRINS ADHESION BINDING PROBES CYCLOADDITION
DOI	10.1021/ja410086d
英文摘要	Metabolic labeling of glycans with chemical reproters (e.g., alkyne or azide) in conjunction with bioorthogonal chemistry is a powerful tool for imaging glycome; however, this method lacks protein-specificity and therefore is not applicable to imaging glycosylation of a specific protein of interest (POI). Here we report the development of a cis-membrane FRET-based methodology that allows protein-specific imaging of glycans on live cells. We exploit metabolic glycan labeling in conjunction with site-specific protein labeling to simultaneously install a FRET acceptor and a donor onto the glycans and the extracellular terminal of the protein of interest, respectively. The intramolecular donor-acceptor distance for the POI falls within the range for effective FRET, whereas the intermolecular FRET is disfavored since the excess acceptors on other proteins are distant from the donor. We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin alpha(x)beta(2), epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for beta(2) integrin activation. Our methodology should enable the live-cell studies on how glycosylation regulates the functions and dynamics of various cell-surface proteins.; Chemistry, Multidisciplinary; SCI(E); EI; 9; ARTICLE; xingchen@pku.edu.cn; 2; 679-687; 136
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/216438]
专题	化学与分子工程学院生命科学学院
推荐引用方式 GB/T 7714	Lin, Wei,Du, Yifei,Zhu, Yuntao,et al. A Cis-Membrane FRET-Based Method for Protein-Specific Imaging of Cell-Surface Glycans[J]. journal of the american chemical society,2014.
APA	Lin, Wei,Du, Yifei,Zhu, Yuntao,&Chen, Xing.(2014).A Cis-Membrane FRET-Based Method for Protein-Specific Imaging of Cell-Surface Glycans.journal of the american chemical society.
MLA	Lin, Wei,et al."A Cis-Membrane FRET-Based Method for Protein-Specific Imaging of Cell-Surface Glycans".journal of the american chemical society (2014).