β-elimination

An unexpected artefact in proteome analysis

Ben Herbert*, Femia Hopwood, David Oxley, John McCarthy, Matt Laver, J. Grinyer, A. Goodall, Keith Williams, Annalisa Castagna, Pier Giorgio Righetti

*Corresponding author for this work

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

Two persistent myths, ingrained in the electrophoretic literature of the last thirty years, namely carbamylation and deamidation, have been recently dispelled (Herbert et al., J. Proteome Res. 2002, in press). We report here, for the first time, a noxious and unexpected artefact in proteome analysis: β-elimination (or desulfuration), which results on the loss of an H2S group (34 Da) from cysteine (Cys) residues for protein focusing in the alkaline pH region. With such an elimination event, a dehydro alanine residue is generated at the Cys site. In turn, the presence of a double bond in this position elicits lysis of the peptide bond, generating a number of peptides of fairly large size from an intact protein. The first process seems to be favored by the electric field, probably due to the continuous harvesting of the SH- anion produced. The only remedy found to this noxious degradation pathway is the reduction and alkylation of all Cys residues prior to their exposure to the electric field. Alkylation appears to substantially reduce both β-elimination and the subsequent amido bond lysis.

Original languageEnglish
Pages (from-to)826-831
Number of pages6
JournalProteomics
Volume3
Issue number6
DOIs
Publication statusPublished - 1 Jun 2003

Keywords

  • β-elimination
  • Carbamylation
  • Proteome analysis
  • Two-dimensional maps

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    Herbert, B., Hopwood, F., Oxley, D., McCarthy, J., Laver, M., Grinyer, J., ... Righetti, P. G. (2003). β-elimination: An unexpected artefact in proteome analysis. Proteomics, 3(6), 826-831. https://doi.org/10.1002/pmic.200300414