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

5,077 Views

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10:36 min

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June 15, 2021

DOI:

10.3791/61983-v

DOI:
10.3791/61983-v

•

10:36 min
June 15, 2021

•

7 Views
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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

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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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Kirsch, Z. J., Arden, B. G., Vachet, R. W., Limpikirati, P. Covalent Labeling with Diethylpyrocarbonate for Studying Protein Higher-Order Structure by Mass Spectrometry. J. Vis. Exp. (172), e61983, doi:10.3791/61983 (2021).

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