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研究膜蛋白的构象动力学原位网站的荧光标记
JoVE Journal
Biology
This content is Free Access.
JoVE Journal Biology
Examining the Conformational Dynamics of Membrane Proteins in situ with Site-directed Fluorescence Labeling
DOI:

11:55 min

May 29, 2011

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Chapters

  • 00:05Title
  • 01:32Protein Expression Cysteine Mutation and mRNA Synthesis
  • 02:07Oocyte Removal
  • 03:36Oocyte Defoculation and mRNA Injection
  • 05:05Site-specific Fluorescence Labeling
  • 06:07Electrophysiology
  • 07:26Anisotropy Measurements and Determination of Distance Constraints
  • 09:17Relative Movement of Protein Subunits
  • 10:07Results: Current and Fluorescence Changes with Site Specific Fluorophore Labelings
  • 11:38Conclusion

Summary

Automatic Translation

我们将介绍一种方法,措施以及具体站点的使用对单个细胞的荧光的构象变化的分析膜蛋白的离子转运的动力学。这种技术是适应离子通道,转运和离子泵并可以被用来确定蛋白亚基之间的距离限制。

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