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相关概念视频

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

2.1K
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...
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¹H NMR of Conformationally Flexible Molecules: Temporal Resolution00:52

¹H NMR of Conformationally Flexible Molecules: Temporal Resolution

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At room temperature, the chair conformer of cyclohexane undergoes rapid ring flipping between two equivalent chair conformers at a rate of approximately 105 times per second. These two chair conformers are in equilibrium. The rapid ring flipping results in the interconversion of the axial proton to an equatorial proton and an equatorial to the axial proton. Such interconversions are too rapid and cannot be detected on the NMR timescale. Hence, the NMR spectrometer cannot distinguish between the...
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相关实验视频

Updated: May 27, 2025

Using Three-color Single-molecule FRET to Study the Correlation of Protein Interactions
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Using Three-color Single-molecule FRET to Study the Correlation of Protein Interactions

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在单分子FRET中扩散具有形态动态的分子的最大概率分析

Irina V Gopich1, John M Louis1, Hoi Sung Chung1

  • 1Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892, United States.

The journal of physical chemistry. B
|February 18, 2025
PubMed
概括

本研究介绍了单分子弗斯特共振能量转移 (FRET) 实验的增强最大概率分析 (burstML),准确地描述了分子动力学和扩散. 改进的方法提供了强大的分析,即使在具有挑战性的实验条件,如高噪音和各种分子状态.

科学领域:

  • 生物物理学的生物物理.
  • 单分子光谱学 单分子光谱学
  • 蛋白质动力学 蛋白质动力学

背景情况:

  • 分析单分子Förster共振能量转移 (FRET) 数据是复杂的,因为分子过渡,扩散和背景噪声.
  • 现有的方法往往通过排除翻译扩散来简化分析,在非理想条件下限制准确性.

研究的目的:

  • 扩展光子爆发 (burstML) 的最大概率分析,用于单个扩散分子的综合分析.
  • 整合用于同时分析形态动力学和转化扩散的方法.
  • 在各种实验挑战下提供分子参数的准确表征.

主要方法:

  • 开发了一种扩展的burstML方法,将扩散和结构动态分析结合起来.
  • 综合现有的仅扩散和仅动态分析方法.
  • 在模拟数据上验证的性能和与除扩散的colorML方法相比.

主要成果:

  • 增强的burstML方法准确地确定了亮度,扩散时间,FRET效率和过渡率.
  • 即使在快速/慢速过渡,高背景噪音和不平等状态亮度/扩散度的情况下,也实现了强大的分析.
  • 在实验条件偏离理想时,与colorML相比,证明了更高的准确性.

更多相关视频

Visualizing the Conformational Dynamics of Membrane Receptors Using Single-Molecule FRET
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Image Processing Protocol for the Analysis of the Diffusion and Cluster Size of Membrane Receptors by Fluorescence Microscopy
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相关实验视频

Last Updated: May 27, 2025

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Published on: January 30, 2018

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Visualizing the Conformational Dynamics of Membrane Receptors Using Single-Molecule FRET
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Visualizing the Conformational Dynamics of Membrane Receptors Using Single-Molecule FRET

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Image Processing Protocol for the Analysis of the Diffusion and Cluster Size of Membrane Receptors by Fluorescence Microscopy
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结论:

  • 扩展爆发ML方法提供了一个更全面,更准确的单分子FRET数据分析与扩散排除方法相比.
  • 这种方法适用于更广泛的实验场景,包括蛋白质折叠研究.
  • 提供了关于在具有挑战性的条件下分子行为的宝贵见解.