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光谱分析 TRPV1 的纯化与重构

JoVE Journal
Bioquímica

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JoVE Journal Bioquímica

Purification and Reconstitution of TRPV1 for Spectroscopic Analysis

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光谱分析 TRPV1 的纯化与重构

DOI:

10.3791/57796-v

•

11:53 min

•

July 03, 2018

•

Francisco J. Sierra-Valdez, Richard A. Stein, Phanindra Velissety³, Valeria Vasquez, Julio F. Cordero-Morales

¹Department of Physiology,University of Tennessee Health Science Center, ²Department of Molecular Physiology and Biophysics,Vanderbilt University Medical Center, ³CuriRX, Inc.

Capítulos

00:04Título
00:54TRPV1 Mutagenesis
02:19Generating the Recombinant Bacmid and Baculovirus for Protein Expression
05:14eTRPV1 and Single-cysteine Mutant Purification
07:18eTRPV1 Single-cysteine Mutant Site-directed Spin Labeling
08:12eTRPV1 Spin-labeled Single Cysteine Mutant Reconstitution
09:30Results: Characterization of the Minimal Cysteine-less TRPV1 Construct and Single-cysteine Mutants used for Spectroscopic Analysis
11:08Conclusion

Summary

Tadução automática

本文介绍了获得洗涤剂-可溶性 TRPV1 的生化量的具体方法, 用于光谱分析。这些联合协议提供了生物化学和生物物理工具, 可用于促进膜控制环境中哺乳动物离子通道的结构和功能研究。

Tags

TRPV1 Ion Channel Spectroscopic Analysis Electron Paramagnetic Resonance Double Electron Electron Resonance Conformational Changes Thermal Gating Light-independent Gating PCR Mutagenesis Recombinant Bacmid ETRPV1 Cysteine Mutants

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