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Updated: Mar 3, 2026

Covalent Labeling with Diethylpyrocarbonate for Studying Protein Higher-Order Structure by Mass Spectrometry
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Covalent Protein Labeling at Glutamic Acids.

Pablo Martín-Gago1, Eyad K Fansa2, Michael Winzker1

  • 1Department of Chemical Biology, Max Planck Institute of Molecular Physiology, Otto-Hahn Straße 11, 44227 Dortmund, Germany.

Cell Chemical Biology
|April 25, 2017
PubMed
Summary
This summary is machine-generated.

Researchers developed a new method for selectively labeling carboxylic acids in proteins using isoxazolium salts. This chemical biology approach targets aspartate and glutamate residues, enabling new protein identification and characterization strategies.

Keywords:
PDE6δWRKWoodward's reagent Kcovalent protein inhibitionglutamic acid labelingisoxazolium salts

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Area of Science:

  • Chemical Biology
  • Proteomics
  • Organic Chemistry

Background:

  • Covalent labeling of proteins targets reactive amino acids like cysteine and lysine.
  • Selective chemical modification of less-reactive carboxylic acids (Asp, Glu) remains a challenge in proteomics.

Purpose of the Study:

  • To develop a novel method for selective covalent targeting of carboxylic acids in proteins.
  • To demonstrate the utility of isoxazolium salts for labeling binding site residues.

Main Methods:

  • Incorporation of isoxazolium salts into protein ligands.
  • Utilizing the lipoprotein binding chaperone PDE6δ as a model system.
  • Analysis of covalent adduct formation and stability.

Main Results:

  • Demonstrated selective covalent labeling of carboxylic acids in Asp and Glu residues.
  • Isoxazolium salts, mimicking Woodward's reagent K, enabled targeting of binding site carboxylic acids.
  • Stable covalent adducts formed via enol esters, resistant to nucleophiles.

Conclusions:

  • A new chemical biology tool for selective carboxylic acid targeting in proteomes has been established.
  • This method expands opportunities for protein identification, characterization, and functional studies.
  • The stable covalent bond formation offers robust labeling for complex biological systems.