单分子荧光成像研究TGF-β信号通路下游蛋白Smad7的抑制机理
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- 英文论文题名：Single-molecule imaging reveals inhibitory mechanism of TGF-β down-stream protein Smad7
- 论文作者：李楠 ; 严晓华 ; 何康敏 ; 袁景和 ; 方晓红
- 英文论文作者：Nan Li ; Xiaohua Yan ; Kangmin He ; Jinghe Yuan ; Xiaohong Fang ; Beijing National Laboratory for Molecular Sciences ; Key Laboratory of Molecular Nanostructure and Nanotechnology ; Institute of Chemistry ; Chinese Academy of Sciences
- 年：2016
- 作者机构：中国科学院化学研究所;
- 论文关键词：单分子荧光成像 ; 单分子追踪 ; Smad7 ; 动力学定量分析
- 会议召开时间：2016-07-01
- 会议录名称：中国化学会第30届学术年会摘要集-第二十八分会：化学生物学
- 语种：中文
- 分类号：Q75
- 学会代码：ZGHY
- 会议名称：中国化学会第30届学术年会-第二十八分会 ; 化学生物学
- 会议地点：中国辽宁大连
- 主办单位：中国化学会
- 学会名称：中国化学会
- 页数：1
- 文件大小：224k
- 原文格式：D
- 会议级别：全国
摘要
转化生长因子TGF-β信号通路在细胞的组织器官发育、细胞增殖、分化、运动以及凋亡等过程中具有非常重要的调节作用。Smad7作为TGF-β信号的关键负调控因子,能够利用多种分子机制抑制TGF-β信号,从而在TGF-β信号通路相关的疾病的发生与发展中有着非常重要的作用。利用传统的生物学方法,如免疫共沉淀和免疫荧光的方法研究Smad7对TGF-β信号的抑制作用,难以观测Smad7在细胞膜上的动态过程和调控机制。我们采用全内反射荧光成像技术,发展并且建立单分子追踪以及统计方法,通过对细胞膜区的EGFP-Smad7进行活细胞单分子荧光成像,我们实时地观察到在TGF-β信号通路中单个Smad7分子上膜的动态行为,并提出Smad7在细胞膜上的抑制机制。
Transforming growth factor β(TGF-β) signaling plays an important role in regulating a broad range of biological processes such as development,growth,differentiation and apoptosis in cells.Smad7 as a negatively regulate factor can antagonize TGF-β signaling via several mechanisms;therefore plays a pivotal role in disease progress.By traditional biochemical assays,such as co-immunoprecipitation or immunofluorescence,the dynamic process of Smad7 membrane docking and regulation is not clear.By using total internal inflection fluorescence microscopy,we developed and established advanced single-molecule tracking and statistical methods.We focused on the real-time study of the dynamic behavior of down-stream cytoplasmic Smad proteins,Smad7.Using advanced live-cell single-molecule fluorescence microscopy to image EGFP-labeled Smad7 on the cell membrane,we observed the dynamics of single Smad7 docking to the cell membrane for TGF-β signaling in real-time,and proposed a model depicting the inhibition role of Smad7 at the cell membrane.
Transforming growth factor β(TGF-β) signaling plays an important role in regulating a broad range of biological processes such as development,growth,differentiation and apoptosis in cells.Smad7 as a negatively regulate factor can antagonize TGF-β signaling via several mechanisms;therefore plays a pivotal role in disease progress.By traditional biochemical assays,such as co-immunoprecipitation or immunofluorescence,the dynamic process of Smad7 membrane docking and regulation is not clear.By using total internal inflection fluorescence microscopy,we developed and established advanced single-molecule tracking and statistical methods.We focused on the real-time study of the dynamic behavior of down-stream cytoplasmic Smad proteins,Smad7.Using advanced live-cell single-molecule fluorescence microscopy to image EGFP-labeled Smad7 on the cell membrane,we observed the dynamics of single Smad7 docking to the cell membrane for TGF-β signaling in real-time,and proposed a model depicting the inhibition role of Smad7 at the cell membrane.
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