Quantitative Characterization of the Membrane Dynamics of Newly Delivered TGF-beta Receptors by Single-Molecule Imaging
文献类型：期刊论文
| 作者 | Zhang, Mingliang1,2,3; Zhang, Zhen3,4; He, Kangmin1,2,3; Wu, Jimin1,2; Li, Nan3,4; Zhao, Rong3,4; Yuan, Jinghe3,4; Xiao, Han1,2; Zhang, Youyi1,2; Fang, Xiaohong3,4 |
| 刊名 | ANALYTICAL CHEMISTRY
 |
| 出版日期 | 2018-04-03 |
| 卷号 | 90期号:7页码:4282-4287 |
| ISSN号 | 0003-2700 |
| DOI | 10.1021/acs.analchem.7b03448 |
| 英文摘要 | The dynamics and stoichiometry of receptors newly delivered on the plasma membrane play a vital role in cell signal transduction, yet knowledge of this process is limited because of the lack of suitable analytical methods. Here we developed a new strategy that combines single-molecule imaging (SMI) and fluorescence recovery after photobleaching (FRAP), named FRAP-SMI, to monitor and quantify individual newly delivered and inserted transmembrane receptors on plasma membranes of living cells. Transforming-growth-factor-beta type II receptor (T beta RII), a typical serine/threoninekinase receptor, was studied with this method. We first eliminated the fluorescence signals from the pre-existing EGFP-labeled T beta RII molecules on the plasma membrane, and then we recorded the individual newly appeared T beta RII-GFP by total-internal-reflection fluorescence imaging. The fluorescence-intensity distributions, photobleaching steps, and diffusion rates of the single T beta RII-GFP molecules were analyzed. We reported, for the first time, that T beta RII was transported to the plasma membrane mainly in the monomeric form in both resting and TGF-beta lstimulated cells. This strongly supported our former discovery that T beta RII could exist as a monomer on the cell membrane, We also found that ligand stimulation resulted in enhanced delivery rates and prolonged membrane association times for the T beta RII molecules. On the basis of these observations, we proposed a mechanism of TGF-beta l-induced T beta RII dimerization for receptor activation. Our method provides a useful tool for the real-time quantification of the spatial arrangement, mobility, and oligomerization of cell-surface proteins in living cells, thus providing a better understanding of cell signaling. |
| 语种 | 英语 |
| 出版者 | AMER CHEMICAL SOC |
| WOS记录号 | WOS:000429385800009 |
| 源URL | [http://ir.iccas.ac.cn/handle/121111/46261]  |
| 专题 | 中国科学院化学研究所 |
| 通讯作者 | Zhang, Youyi; Fang, Xiaohong |
| 作者单位 | 1.Peking Univ, Hosp 3, Inst Vasc Med,Beijing Key Lab Cardiovasc Receptor, Minist Hlth,Key Lab Cardiovasc Mol Biol & Regulat, Beijing 100191, Peoples R China 2.Peking Univ, Acad Adv Interdisciplinary Studies, Beijing 100191, Peoples R China 3.Chinese Acad Sci, CAS Res Educ Ctr Excellence Mol Sci, Inst Chem, Key Lab Mol Nanostruct & Nanotechnol, Beijing 100190, Peoples R China 4.Univ Chinese Acad Sci, Beijing 100049, Peoples R China |
| 推荐引用方式 GB/T 7714 | Zhang, Mingliang,Zhang, Zhen,He, Kangmin,et al. Quantitative Characterization of the Membrane Dynamics of Newly Delivered TGF-beta Receptors by Single-Molecule Imaging[J]. ANALYTICAL CHEMISTRY,2018,90(7):4282-4287. |
| APA | Zhang, Mingliang.,Zhang, Zhen.,He, Kangmin.,Wu, Jimin.,Li, Nan.,...&Fang, Xiaohong.(2018).Quantitative Characterization of the Membrane Dynamics of Newly Delivered TGF-beta Receptors by Single-Molecule Imaging.ANALYTICAL CHEMISTRY,90(7),4282-4287. |
| MLA | Zhang, Mingliang,et al."Quantitative Characterization of the Membrane Dynamics of Newly Delivered TGF-beta Receptors by Single-Molecule Imaging".ANALYTICAL CHEMISTRY 90.7(2018):4282-4287. |
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来源：化学研究所
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