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Journal
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Biochemistry
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利用体外荧光共振能量转移研究毫秒时间尺度蛋白质配合物的动力学
JoVE Journal
Biochemistry
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JoVE Journal
Biochemistry
Using In Vitro Fluorescence Resonance Energy Transfer to Study the Dynamics Of Protein Complexes at a Millisecond Time Scale
Please note that all translations are automatically generated.
Click here for the English version.
利用体外荧光共振能量转移研究毫秒时间尺度蛋白质配合物的动力学
DOI:
10.3791/59038-v
•
10:50 min
•
March 14, 2019
•
Melaku Garsamo
3
,
Yun Zhou
3
,
Xing Liu
3
1
Department of Biochemistry
,
Purdue University
,
2
Department of Botany and Plant Pathology
,
Purdue University
,
3
Center for Plant Biology
,
Purdue University
Chapters
00:04
Title
00:33
Designing the FRET Assay
01:12
Preparation of Cul1
AMC
•Rbx1, the FRET Donor Protein
04:42
Preparation of
FlAsH
Cand1, the FRET Acceptor Protein
05:18
Testing and Confirmation of the FRET Assay
06:08
Measuring the Association Rate Constant
07:37
Measuring the Dissociation Rate Constant of Cul1•Cand1 in the Presence of Skp•F-box Protein
08:33
Results: Dynamics Of Protein Complexes
10:30
Conclusion
Summary
Automatic Translation
English (Original)
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Automatic Translation
蛋白质-蛋白质相互作用对生物系统至关重要, 对结合动力学的研究为蛋白质复合物的动力学和功能提供了见解。我们描述了一种利用荧光共振能量转移和停止流动技术量化蛋白质复合物动力学参数的方法。
Tags
FRET
Protein Complex
Kinetics
Association
Dissociation
COL1
CAND1
7-amino-4-methylcoumarin
FlASH
Cell Lysis
Sonication
Centrifugation
Glutathione Beads
Elution
Thrombin
Buffer A
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