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Disulfide Connectivity Analysis of Peptides Bearing Two Intramolecular Disulfide Bonds Using MALDI In-Source Decay

  • Philippe Massonnet
  • Jean R. N. Haler
  • Gregory Upert
  • Nicolas Smargiasso
  • Gilles Mourier
  • Nicolas Gilles
  • Loïc Quinton
  • Edwin De Pauw
Research Article

Abstract

Disulfide connectivity in peptides bearing at least two intramolecular disulfide bonds is highly important for the structure and the biological activity of the peptides. In that context, analytical strategies allowing a characterization of the cysteine pairing are of prime interest for chemists, biochemists, and biologists. For that purpose, this study evaluates the potential of MALDI in-source decay (ISD) for characterizing cysteine pairs through the systematic analysis of identical peptides bearing two disulfide bonds, but not the same cysteine connectivity. Three different matrices have been tested in positive and/or in negative mode (1,5-DAN, 2-AB and 2-AA). As MALDI-ISD is known to partially reduce disulfide bonds, the data analysis of this study rests firstly on the deconvolution of the isotope pattern of the parent ions. Moreover, data analysis is also based on the formed fragment ions and their signal intensities. Results from MS/MS-experiments (MALDI-ISD-MS/MS) constitute the last reference for data interpretation. Owing to the combined use of different ISD-promoting matrices, cysteine connectivity identification could be performed on the considered peptides.

Graphical Abstract

Keywords

Mass spectrometry Peptide Disulfide bonds MALDI ISD Isomers 

Notes

Acknowledgments

The authors thank the FRS-FNRS for the financial support (FRIA and instrumentation), the Fonds Européen de développement regional (FEDER), the Walloon region, and the European commission (F.P. 7 VENOMICS project) for financial support.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

13361_2018_2022_MOESM1_ESM.pdf (1.1 mb)
ESM 1 (PDF 1.08 mb)

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Copyright information

© American Society for Mass Spectrometry 2018

Authors and Affiliations

  1. 1.Mass Spectrometry Laboratory, MolSys Research UnitUniversity of LiègeLiegeBelgium
  2. 2.Commissariat à l’Energie Atomique, DRF/SIMOPROGif sur YvetteFrance

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