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Förster Resonance Energy Transfer Mapping: A New Methodology to Elucidate Global Structural Features
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Biochemistry
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JoVE Journal Biochemistry
Förster Resonance Energy Transfer Mapping: A New Methodology to Elucidate Global Structural Features

Förster Resonance Energy Transfer Mapping: A New Methodology to Elucidate Global Structural Features

DOI:
10.3791/63433-v

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07:09 min

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March 16, 2022

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, ,
1Molecular Biology and Biochemistry Department,Wesleyan University, 2Molecular Biophysics Program,Wesleyan University

Chapters

  • 00:04Introduction
  • 00:59Selection of Labeling Sites
  • 01:29Determination of the R0 Values
  • 03:04Performing FRET Spectral Measurements
  • 04:09Mapping the Distances
  • 04:46Results: Förster Resonance Energy Transfer Mapping of the Preprotein Binding Site
  • 06:30Conclusion

Summary

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The study details the methodology of FRET mapping including the selection of labeling sites, choice of dyes, acquisition, and data analysis. This methodology is effective at determining binding sites, conformational changes, and dynamic motions in protein systems and is most useful if performed in conjunction with existing 3-D structural information.

Tags

Keywords Extracted From The Given Text:FRET MappingLigand Binding SitesSubunit OrientationConformational ChangesDynamic MotionsBiomolecular SystemLabeling SitesR0 ValuesDonor DyeAcceptor DyeSpectrofluorometerEmission ScanExcitation Wavelength

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