相关实验视频

Updated: Jul 18, 2025

Purification and Reconstitution of TRPV1 for Spectroscopic Analysis

Purification and Reconstitution of TRPV1 for Spectroscopic Analysis

Published on: July 3, 2018

使用衍射X射线追踪方法实时观察TRPV1的素诱导的细胞内域动力学

Kazuhiro Mio^1,2, Tatsunari Ohkubo^1,2, Daisuke Sasaki³

¹AIST-UTokyo Advanced Operando-Measurement Technology Open Innovation Laboratory (OPERANDO-OIL), National Institute of Advanced Industrial Science and Technology (AIST), 6-2-3 Kashiwanoha, Chiba 277-0882, Japan.

|August 25, 2023

概括

分散X射线追踪揭示了TRPV1通道的高频动态. 素诱导反时钟方向的C端运动,而AMG9810则导致时钟方向扭转,阐明了TRPV1激活机制.

科学领域:

分子生物学分子生物学
生物物理学的生物物理.
结构生物学是结构生物学.

背景情况:

短暂受体潜在化物1型 (TRPV1) 是一个关键的离子通道,参与疼痛和温度感觉.
虽然TRPV1的结构已知,但由于其复杂性,其关机制仍然不清楚.
了解TRPV1的动态是开发新型止痛药的关键.

研究的目的:

为了研究TRPV1通道的细胞内域动态.
阐明控制TRPV1通道封闭的高频运动.
区分激动剂和对抗剂对TRPV1动态的影响.

主要方法:

使用金纳米晶体的衍射X射线追踪 (DXT) 来捕捉分子内运动.
在高 (100微秒/) 和低 (12.5毫秒/) 速率记录的衍射数据.
应用终身过来分析高速C端波动.

主要成果:

素结合导致TRPV1 C端在高频率下反时钟旋转偏差.
在N端观察到与素独立的内在的反时钟方向扭动运动.
反对者AMG9810诱导了顺时针旋转的运动,与素的影响相反.

结论:

DXT成功捕获了高频TRPV1动态,揭示了明显的运动模式.
素和AMG9810对TRPV1旋转动态产生相反的影响.
这些发现为TRPV1通道的激活和调制机制提供了新的见解.

关键词:

在 TRP 频道中,可以使用 TRP 频道.在 TRPV1 中.形状变化动态的变化动态分散的X射线追踪技术.单个分子分析分析

更多相关视频

Controllable Ion Channel Expression through Inducible Transient Transfection

Controllable Ion Channel Expression through Inducible Transient Transfection

Published on: February 17, 2017

Single-Cell Calcium Imaging for Studying the Activation of Calcium Ion Channels

Single-Cell Calcium Imaging for Studying the Activation of Calcium Ion Channels

Published on: December 13, 2024

相关概念视频

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...