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

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

10,484 Views

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11:24 min

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May 13, 2017

DOI:

10.3791/55623-v

DOI:
10.3791/55623-v

•

11:24 min
May 13, 2017

•

4 Views
, , , , ,
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

Summary

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A protocol for high-precision FRET experiments at the single molecule level is presented here. Additionally, this methodology can be used to identify three conformational states in the ligand-binding domain of the N-methyl-D-aspartate (NMDA) receptor. Determining precise distances is the first step towards building structural models based on FRET experiments.

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

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Cite this Article

Ma, J., Yanez-Orozco, I. S., Rezaei Adariani, S., Dolino, D., Jayaraman, V., Sanabria, H. High Precision FRET at Single-molecule Level for Biomolecule Structure Determination. J. Vis. Exp. (123), e55623, doi:10.3791/55623 (2017).

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