相关实验视频

Updated: Jun 29, 2025

Visualization and Quantification of TGFβ/BMP/SMAD Signaling under Different Fluid Shear Stress Conditions using Proximity-Ligation-Assay

Visualization and Quantification of TGFβ/BMP/SMAD Signaling under Different Fluid Shear Stress Conditions using Proximity-Ligation-Assay

Published on: September 14, 2021

基于TurboID近距离标记技术的转录因子Smad3相互作用蛋白的研究

Biao Yan¹, Ting Zeng², Xiaoshan Liu³

¹School of Basic Medical Sciences, Xianning Medical College, Hubei University of Science and Technology, Xianning 437000, Hubei, China.

|March 27, 2024

概括

研究人员使用TurboID识别与Smad3相互作用的蛋白质,Smad3是TGF-β信号传递的关键参与者. 他们发现hnRNPM与Smad3相互作用,Smad3影响IL-6的表达,为糖尿病病和炎症提供了洞察力.

科学领域:

分子生物学分子生物学
细胞生物学细胞生物学
生物化学生物化学

背景情况:

转化生长因子-β (TGF-β) /Smad3信号通路与各种疾病有关,包括糖尿病病和炎症.
识别相互作用的蛋白质对于理解Smad3的调节网络和功能至关重要.

研究的目的:

通过识别Smad3相互作用蛋白来研究TGF-β/Smad3信号通路的功能.
为蛋白质相互作用研究构建表达Smad3的稳定细胞系.

主要方法:

使用lentiviral感染构建一个稳定的Smad3-转染细胞系.
使用TurboID进行蛋白质标签,然后进行无标签质谱测量以识别相互作用的蛋白质.
免疫沉 (IP) 和免疫光测试用于验证相互作用和基因表达效应.

主要成果:

质谱学确定了491种可能与Smad3.3相互作用的蛋白质.
异质核核核糖核蛋白M (hnRNPM) 被确定为一种相互作用蛋白.
发现Smad3促进了hnRNPM的表达,并随后抑制了Interleukin-6 (IL-6) 的表达.

结论:

Smad3 与 hnRNPM 相互作用,影响其表达.
Smad3-hnRNPM相互作用对IL-6具有抑制作用,这表明它在炎症过程中起作用.
这项研究为进一步研究Smad3的功能及其在疾病环境中的调控网络提供了基础.

关键词:

生物素标签的标签这真是太疯狂了这是一个TurboIDID.U251 是一个U251的元素.在 hnRNPM 里面.

更多相关视频

Detection of Heterodimerization of Protein Isoforms Using an in Situ Proximity Ligation Assay

Detection of Heterodimerization of Protein Isoforms Using an in Situ Proximity Ligation Assay

Published on: October 20, 2018

Live Cell Imaging of the TGF- β/Smad3 Signaling Pathway In Vitro and In Vivo Using an Adenovirus Reporter System

Live Cell Imaging of the TGF- β/Smad3 Signaling Pathway In Vitro and In Vivo Using an Adenovirus Reporter System

Published on: July 30, 2018

相关概念视频

TGF - β Signaling Pathway

TGF - β Signaling Pathway

The TGF-β signaling pathway regulates cell growth, differentiation, adhesion, motility, and development. TGF-β ligands that induce TGF-β signaling are synthesized in their latent form. Several proteases or cell surface receptors such as integrins act upon the latent form, releasing the active ligand. There are three types of mammalian TGF-βs: (TGF-β1, TGF-β2, and TGF-β3) that bind as homodimers or heterodimers to TGF-β receptors. The TGF-β receptors...

Protein Dynamics in Living Cells

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...