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Luminescence Resonance Energy Transfer to Study Conformational Changes in Membrane Proteins Expressed in Mammalian Cells
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Bioengineering
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JoVE Journal Bioengineering
Luminescence Resonance Energy Transfer to Study Conformational Changes in Membrane Proteins Expressed in Mammalian Cells

Luminescence Resonance Energy Transfer to Study Conformational Changes in Membrane Proteins Expressed in Mammalian Cells

DOI:
10.3791/51895-v

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08:31 min

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September 16, 2014

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, ,
1Center for Membrane Biology, Department of Biochemistry and Molecular Biology,University of Texas Health Science Center at Houston

Chapters

  • 00:05Title
  • 01:29Protein Tagging Site and Fluorophore Selection
  • 03:04Luminescence Resonance Energy Transfer (LRET) Setup
  • 05:00Data Analysis
  • 06:58Results: Representative LRET Measurements
  • 07:57Conclusion

Summary

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We describe here an improved Luminescence Resonance Energy Transfer (LRET) method where we introduce a protease cleavage site between the donor and acceptor fluorophore sites. This modification allows us to obtain specific LRET signals arising from membrane proteins of interest, allowing for the study of membrane proteins without protein purification.

Tags

Luminescence Resonance Energy TransferLRETMembrane ProteinsConformational ChangesLanthanideTerbiumDonor-acceptor PairProtease Cleavage SiteMammalian Cells

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