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使用单分子FRET可视化膜受体的构象动力学
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Visualizing the Conformational Dynamics of Membrane Receptors Using Single-Molecule FRET
DOI:

10:59 min

August 17, 2022

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Chapters

  • 00:04Introduction
  • 01:04Slide and Coverslip Cleaning
  • 03:04mGluR2 Expression with Incorporated Unnatural Amino Acid, Fluorescent Labeling, and Extraction
  • 05:13Single-Molecule Flow Chamber Assembly and Functionalization
  • 06:23Microscope Setup and smFRET Data Acquisition
  • 07:47Data Analysis
  • 08:39Results: Analyzing Conformational Dynamics of Membrane Receptors Proteins
  • 10:11Conclusion

Summary

Automatic Translation

本研究提出了一种详细的程序,通过非天然氨基酸(UAA)掺入 使用 位点特异性标记对G蛋白偶联受体(GPCR)进行单分子荧光共振能量转移(smFRET)实验。该协议为 smFRET 样品制备、实验和数据分析提供了分步指南。

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