Combination of Capillary Zone Electrophoresis-Mass Spectrometry, Ion Mobility-Mass Spectrometry, and Theoretical Calculations for Cysteine Connectivity Identification in Peptides Bearing Two Intramolecular Disulfide Bonds

Cedric Delvaux*, Philippe Massonnet, Christopher Kune, Jean R. N. Haler, Gregory Upert, Gilles Mourier, Nicolas Gilles, Loic Quinton, Edwin De Pauw, Johann Far

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

9 Citations (Web of Science)

Abstract

Disulfide bonds between cysteine residues are commonly involved in the stability of numerous peptides and proteins and are crucial for providing biological activities. In such peptides, the appropriate cysteine connectivity ensures the proper conformation allowing an efficient binding to their molecular targets. Disulfide bond connectivity characterization is still challenging and is a critical issue in the analysis of structured peptides/proteins targeting pharmaceutical or pharmacological utilizations. This study describes the development of new and fast gas-phase and in-solution electrophoretic methods coupled to mass spectrometry to characterize the cysteine connectivity of disulfide bonds. For this purpose, disulfide isomers of three peptides bearing two intramolecular disulfide bonds but different cysteine connectivity have been investigated. Capillary zone electrophoresis and ion mobility both coupled to mass spectrometry were used to perform the separation in both aqueous and gas phases, respectively. The separation efficiency of each technique has been critically evaluated and compared. Finally, theoretical calculations were performed to support and explain the experimental data based on the predicted physicochemical properties of the different peptides.

Original languageEnglish
Pages (from-to)2425-2434
Number of pages10
JournalAnalytical Chemistry
Volume92
Issue number3
DOIs
Publication statusPublished - 4 Feb 2020

Keywords

  • COLLISION-INDUCED DISSOCIATION
  • ELECTRON-TRANSFER DISSOCIATION
  • PROTEIN STABILITY
  • IONIZABLE GROUPS
  • PK VALUES
  • CONOTOXIN
  • ISOMERS
  • TOOL

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