Distinguishing between phosphorylated and nonphosphorylated peptides with ion mobility-mass spectrometry

Brandon T. Ruotolo, Guido F. Verbeck IV, Lisa M. Thomson, Amina S. Woods, Kent J. Gillig, David H. Russell

Research output: Contribution to journalArticle

Abstract

Mass spectrometry has become an indispensable tool in identifying post-translationally modified proteins, but multiple peptide mass-mapping/peptide-sequencing experiments are required to answer questions involving the site and type of modification present. Here, we apply ion mobility-mass spectrometry (IM-MS), a high-throughput analysis method having high selectivity and sensitivity, to the challenge of identifying phosphorylated peptides. Ion mobility separation is based on the collision cross-section of the ion. Phosphorylation can result in a conformational change in gas-phase peptide ions, which can be detected by IM. To demonstrate this point, a peptide mixture containing a variety of peptide sequences is examined with IM-MS and molecular dynamics calculations. During the course of these studies, two classes of phosphopeptide were identified: (i) phosphorylated peptide ions that have conformers that differ from the nonphosphorylated ion and (ii) phosphorylated peptide ions that have conformations that are very similar to the nonphosphorylated peptide. The utility of IM-MS peptide mass mapping for identifying both types of phosphorylated peptides is discussed.

Original languageEnglish (US)
Pages (from-to)303-306
Number of pages4
JournalJournal of Proteome Research
Volume1
Issue number4
DOIs
StatePublished - Jul 2002
Externally publishedYes

Keywords

  • Conformation
  • High-throughput
  • MALDI
  • Phosphopeptides

ASJC Scopus subject areas

  • Genetics
  • Biotechnology
  • Biochemistry

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    Ruotolo, B. T., Verbeck IV, G. F., Thomson, L. M., Woods, A. S., Gillig, K. J., & Russell, D. H. (2002). Distinguishing between phosphorylated and nonphosphorylated peptides with ion mobility-mass spectrometry. Journal of Proteome Research, 1(4), 303-306. https://doi.org/10.1021/pr025516r