Solid-phase glycosylation of peptide templates and on-bead MAS-NMR analysis: Perspectives for glycopeptide libraries

Charlotte Held Gotfredsen, Koen M. Halkes, Jens Øllgaard Duus, Morten Meldal, Les P. Miranda, Morten Grøtli

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

The efficient solid-phase glycosylation of amino acid side chains (serine (Ser), threonine (Thr), and tyrosine (Tyr)) in peptides was demonstrated with a variety of glycosyl trichloroacetimidate donors in high yields and purities. A novel photolabile linker, with no chiral centre, was introduced to facilitate analysis by both matrix-assisted laser desorption ionisation time of flight (MALDI-TOF) mass spectrometry and nanoprobe magic angle spinning (MAS) NMR spectroscopy. Product analysis by nanoprobe MAS NMR spectroscopy, LC-MS and MALDI-TOF mass spectrometry of the glycosylation reactions indicated that the reactivity order of the hydroxy side-chain functions of amino acids in peptides on the solid-phase was Tyr > Ser > Thr. The nearly quantitative glycosylation yields and the efficient on-bead product analysis by nanoprobe MAS NMR spectroscopy have made a truly solid-phase approach for the synthesis and analysis of glycopeptide libraries possible.
Original languageEnglish
JournalChemistry: A European Journal
Volume7
Issue number16
Pages (from-to)3584-3591
ISSN0947-6539
DOIs
Publication statusPublished - 2001
Externally publishedYes

Keywords

  • Amino Acid Sequence
  • Carbohydrate Conformation
  • Glycopeptides
  • Glycosylation
  • Indicators and Reagents
  • Models, Molecular
  • Nuclear Magnetic Resonance, Biomolecular
  • Peptides
  • Protein Conformation
  • Serine
  • Threonine
  • Tyrosine
  • 2ZD004190S Threonine
  • 42HK56048U Tyrosine
  • 452VLY9402 Serine
  • Ionization
  • Lasers
  • Mass spectrometry
  • Synthesis (chemical)
  • Amino acids
  • article
  • desorption
  • ionization
  • mass spectrometry
  • nuclear magnetic resonance
  • peptide synthesis
  • protein glycosylation
  • solid phase extraction
  • NMR spectroscopy
  • Photolabile linker
  • Solid-phase synthesis
  • T
  • X

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