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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
Please note that all translations are automatically generated.
Click here for the English version.
研究膜蛋白的构象动力学
原位
网站的荧光标记
DOI:
10.3791/2627-v
•
11:55 min
•
May 29, 2011
•
Ryan Richards
,
Robert E. Dempski
1
Department of Chemistry and Biochemistry
,
Worcester Polytechnic Institute
Chapters
00:05
Title
01:32
Protein Expression Cysteine Mutation and mRNA Synthesis
02:07
Oocyte Removal
03:36
Oocyte Defoculation and mRNA Injection
05:05
Site-specific Fluorescence Labeling
06:07
Electrophysiology
07:26
Anisotropy Measurements and Determination of Distance Constraints
09:17
Relative Movement of Protein Subunits
10:07
Results: Current and Fluorescence Changes with Site Specific Fluorophore Labelings
11:38
Conclusion
Summary
Automatic Translation
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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。
Tags
Conformational Dynamics
Membrane Proteins
Site-directed Fluorescence Labeling
Two Electrode Voltage Clamp Electrophysiology (TEVC)
Ion Transport
Ion Channels
Ion Pumps
Transporters
Site-specific Fluorophore Labeling
Conformational Dynamics
Voltage-clamp Fluorometry
Cysteine Replacement
Site-directed Fluorophore Labeling
Molecular Motion
Distance Constraints
Xenopus Leavis Oocytes
Functional Expression
Fluorescence Signal
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