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Biochemistry

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

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

Article DOI: 10.3791/55623-v • 11:24 min • May 13th, 2017
May 13th, 2017 •
1, 2, 2, 3, 3, 2
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

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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 FRET Single-molecule Level Biomolecule Structure Determination Inter-dye Distance Measurements Multi-parameter Fluorescence Detection FA Protocol F&246;rster Resonance Energy Transfer MFD Mode Photophysical Artifacts Experimental Artifacts Interdye Distance Accuracy Rigid Biomolecules Conformational States Ligand-binding Domain N-methyl-D-aspartate Receptor Activation Of The Receptor Crystallographic Structures Dynamic Biomolecules Distance Restraints Structural Model
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