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

Article DOI: 10.3791/61983-v • 10:36 min • June 15th, 2021
June 15th, 2021 •
1, 1, *1,2, *3
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
* These authors contributed equally

总结概括
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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.

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Covalent Labeling Diethylpyrocarbonate Protein Higher-order Structure Mass Spectrometry Protein Characterization Chemical Reactions Protein Structural Information Mass Increases Residue Level Resolution Proteolytic Digestion Tandem Mass Spectrometry Diethyl Pyrocarbonate (DEPC) Labeling Reagent Mass Spec Detection Electrophilic Molecule Structural Resolution Buffered Solution Micromolar Concentration PH 7.4 MOPS Buffer
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