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Covalent Labeling with Diethylpyrocarbonate for Studying Protein Higher-Order Structure by Mass Spectrometry
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JoVE Journal Chemie
Covalent Labeling with Diethylpyrocarbonate for Studying Protein Higher-Order Structure by Mass Spectrometry

Covalent Labeling with Diethylpyrocarbonate for Studying Protein Higher-Order Structure by Mass Spectrometry

DOI:
10.3791/61983-v

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10:36 min

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June 15, 2021

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, , ,
1Department of Chemistry,University of Massachusetts Amherst, 2Molecular and Cellular Biology Program,University of Massachusetts Amherst, 3Department of Food and Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences,Chulalongkorn University

Summary

Automatische Übersetzung

Automatische Übersetzung

The experimental procedures for performing diethylpyrocarbonate-based covalent labeling with mass spectrometric detection are described. Diethylpyrocarbonate is simply mixed with the protein or protein complex of interest, leading to the modification of solvent accessible amino acid residues. The modified residues can be identified after proteolytic digestion and liquid chromatography/mass spectrometry analysis.

Tags

Covalent LabelingDiethylpyrocarbonateDEPCMass SpectrometryProtein StructureStructural CharacterizationBuffer ExchangeImidazole

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