A Cis-Membrane FRET-Based Method for Protein-Specific Imaging of Cell-Surface Glycans
文献类型：期刊论文
| 作者 | Lin, Wei; Du, Yifei; Zhu, Yuntao; Chen, Xing1 |
| 刊名 | JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
 |
| 出版日期 | 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 |
| WOS记录号 | WOS:000330018600032 |
| 出版者 | AMER CHEMICAL SOC |
| 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. |
入库方式： OAI收割
来源：化学研究所
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