Selenopeptide mapping in a selenium-yeast protein digest by parallel nanoHPLC-ICP-MS and nanoHPLC-electrospray-MS/MS after on-line preconcentration

Pierre Giusti, Dirk Schaumlöffel, Hugues Preud'homme, Joanna Szpunar, Ryszard Lobinski

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

ICP collision cell MS was optimized for the detection and retention-time marking of selenium-containing peptides in nanoHPLC (75 μm column) after on-line 100-fold preconcentration on a capillary (300 μm id) precolumn. The mobile phase composition, gradient and flow rate were chosen to allow electrospray-MS/MS to be successfully run in parallel in identical separation and preconcentration conditions in order to produce two matching sets of chromatograms: an element-specific one and a molecule-specific one. Knowledge of the retention time of a Se-containing peptide of interest allowed efficient data mining in the corresponding ES-MS chromatogram and the identification of minor Se-species. A third chromatogram was run to obtain collision-induced dissociation data for the target peptides. The performance of the method was demonstrated for a comprehensive on-line characterization of a mixture of peptides in a tryptic digest of a Se-containing protein fraction isolated by size-exclusion chromatography from a selenium yeast extract. The method allowed the identification of whole series of Se/S substitutions in individual peptides and, in some cases, sequencing of isomers differing in the position of selenomethionine residues in the amino acid sequence.

Original languageEnglish
Pages (from-to)26-32
Number of pages7
JournalJournal of Analytical Atomic Spectrometry
Volume21
Issue number1
DOIs
Publication statusPublished - 10 Apr 2006
Externally publishedYes

Fingerprint

Fungal Proteins
Selenium
Peptides
Selenomethionine
Size exclusion chromatography
Phase composition
Isomers
Yeast
Data mining
Substitution reactions
Flow rate
Amino Acids
Molecules
Proteins

ASJC Scopus subject areas

  • Spectroscopy

Cite this

Selenopeptide mapping in a selenium-yeast protein digest by parallel nanoHPLC-ICP-MS and nanoHPLC-electrospray-MS/MS after on-line preconcentration. / Giusti, Pierre; Schaumlöffel, Dirk; Preud'homme, Hugues; Szpunar, Joanna; Lobinski, Ryszard.

In: Journal of Analytical Atomic Spectrometry, Vol. 21, No. 1, 10.04.2006, p. 26-32.

Research output: Contribution to journalArticle

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