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单分子水平的高精度FRET用于生物分子结构测定
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Biochemistry
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JoVE Journal Biochemistry
High Precision FRET at Single-molecule Level for Biomolecule Structure Determination
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单分子水平的高精度FRET用于生物分子结构测定

DOI:
10.3791/55623-v

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11:24 min

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May 13, 2017

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, , , , ,
1Department of Chemistry,Clemson University, 2Department of Physics and Astronomy,Clemson University, 3Department of Biochemistry and Molecular Biology, Center for Membrane Biology, Graduate School for Biomedical Sciences,University of Texas Health Science Center

Chapters

  • 00:05Title
  • 01:07Preparation of Materials
  • 02:57Pulsed Interleaved Excitation-Multiparameter Fluorescence Detection (PIE-MFD) Instrument Setup
  • 06:14Burst Integration Fluorescence Lifetime Analysis of Standards
  • 07:34dsDNA and Protein Sample Measurements and Analysis
  • 09:28Results: Inter-dye Distance Determination in Single Molecules
  • 10:54Conclusion

Summary

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本文介绍了单分子水平高精度FRET实验方案。此外，该方法可用于鉴定N-甲基-D-天冬氨酸（NMDA）受体的配体结合结构域中的三种构象状态。确定精确的距离是建立基于FRET实验的结构模型的第一步。

Tags

High Precision FRETSingle-moleculeBiomolecule Structure DeterminationFörster Resonance Energy TransferMultiparameter Fluorescence DetectionInterdye Distance MeasurementNMDA ReceptorConformational StatesStructural ModelDynamic Biomolecules

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