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

doi:10.1021/ja410086d

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	A Cis-Membrane FRET-Based Method for Protein-Specific Imaging of Cell-Surface Glycans
	Lin, Wei ; Du, Yifei ; Zhu, Yuntao ; Chen, Xing 1
刊名	JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
	2014-01-15
卷号	136 期号:2 页码:679-687
ISSN号	0002-7863
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.
语种	英语
出版者	AMER CHEMICAL SOC
WOS记录号	WOS:000330018600032
内容类型	期刊论文
源URL	[http://ir.iccas.ac.cn/handle/121111/50703]
专题	中国科学院化学研究所
通讯作者	Chen, Xing
作者单位	1.Peking Univ, Coll Chem & Mol Engn, Synthet & Funct Biomol Ctr, Beijing Natl Lab Mol Sci,Dept Chem Biol, Beijing 100871, Peoples R China 2.Peking Univ, Peking Tsinghua Ctr Life Sci, Beijing 100871, Peoples R China
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,2014,136(2):679-687.
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,136(2),679-687.
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 136.2(2014):679-687.