Journal
/
Bioquímica
/
Using Three-color Single-molecule FRET to Study the Correlation of Protein Interactions
JoVE Journal
Bioquímica
É necessária uma assinatura da JoVE para visualizar este conteúdo.  Faça login ou comece sua avaliação gratuita.
JoVE Journal Bioquímica
Using Three-color Single-molecule FRET to Study the Correlation of Protein Interactions

Using Three-color Single-molecule FRET to Study the Correlation of Protein Interactions

DOI:
10.3791/56896-v

•

11:22 min

•

January 30, 2018

•

, , ,
1Institute of Physical Chemistry,University of Freiburg, 2Department of Bionanoscience, Kavli Institute of Nanoscience Delft,Delft University of Technology

Capítulos

  • 00:05Título
  • 00:37Total Internal Reflection Fluorescence Microscope (TIRFM) Flow Chamber Assembly
  • 02:15Three-color smFRET Measurement of Hsp90
  • 05:52Data Analysis
  • 09:08Results: Protein Interactions of Hsp90
  • 10:47Conclusion

Summary

Tadução automática

Tadução automática

Here, we present a protocol to obtain three-color smFRET data and its analysis with a 3D ensemble Hidden Markov Model. With this approach, scientists can extract kinetic information from complex protein systems, including cooperativity or correlated interactions.

Tags

Three-color Single-molecule FRETProtein InteractionsCooperativityQuantitative BiologyProtein ComplexesDynamic InteractionsFlow Chamber AssemblyPEG-biotin Functionalized SlideMulti-color Prism TIRF MicroscopeEMCCD Camera Configuration

Article

Embed

ADICIONAR À PLAYLIST

Usage Statistics

Vídeos Relacionados

Fluorescence Anisotropy as a Tool to Study Protein-protein Interactions

Structural Information from Single-molecule FRET Experiments Using the Fast Nano-positioning System

Using Scaffold Liposomes to Reconstitute Lipid-proximal Protein-protein Interactions In Vitro

High Precision FRET at Single-molecule Level for Biomolecule Structure Determination

Biotinylated Cell-penetrating Peptides to Study Intracellular Protein-protein Interactions

Covalent Immobilization of Proteins for the Single Molecule Force Spectroscopy

In Vivo Hydroxyl Radical Protein Footprinting for the Study of Protein Interactions in Caenorhabditis elegans

Making Precise and Accurate Single-Molecule FRET Measurements using the Open-Source smfBox

Optical Tweezers to Study RNA-Protein Interactions in Translation Regulation

Single Molecule Fluorescence Energy Transfer Study of Ribosome Protein Synthesis

Read Article