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Journal / Biochemistry / Measuring Nucleotide Binding to Intact,…
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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

Article DOI: 10.3791/61401-v • 08:33 min • March 11th, 2021
March 11th, 2021 •
1, 1, 1,2
1Department of Physiology, Anatomy and Genetics, University of Oxford, 2Department of Chemistry and Neuroscience Program, Trinity College

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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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Keywords: Nucleotide Binding Membrane Proteins FRET ATP Potassium Channel Diabetes Unroofed Membrane Sonication Microscopy ANAP Fluorescence Imaging
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