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JoVE Journal

Bioengineering

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

11.9K Views

08:31 min

September 16th, 2014

September 16th, 2014

11,924 Views

0:05

Title

1:29

Protein Tagging Site and Fluorophore Selection

3:04

Luminescence Resonance Energy Transfer (LRET) Setup

5:00

Data Analysis

6:58

Results: Representative LRET Measurements

7:57

Conclusion

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.

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