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Measuring Nucleotide Binding to Intact, Functional Membrane Proteins in Real Time
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Biochemistry
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JoVE Journal Biochemistry
Measuring Nucleotide Binding to Intact, Functional Membrane Proteins in Real Time

Measuring Nucleotide Binding to Intact, Functional Membrane Proteins in Real Time

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1,706 Views

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08:33 min

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March 11, 2021

DOI:

10.3791/61401-v

DOI:
10.3791/61401-v

•

08:33 min
March 11, 2021

•

1 Views
, ,
1Department of Physiology, Anatomy and Genetics,University of Oxford, 2Department of Chemistry and Neuroscience Program,Trinity College

Summary

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This protocol presents a method for measuring adenine nucleotide binding to receptors in real time in a cellular environment. Binding is measured as Förster resonance energy transfer (FRET) between trinitrophenyl nucleotide derivatives and protein labeled with a non-canonical, fluorescent amino acid.

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Cite this Article

Usher, S. G., Ashcroft, F. M., Puljung, M. C. Measuring Nucleotide Binding to Intact, Functional Membrane Proteins in Real Time. J. Vis. Exp. (169), e61401, doi:10.3791/61401 (2021).

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