title: Solid-phase glycosylation of peptide templates and on-bead MAS-NMR analysis: perspectives for glycopeptide libraries.
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.
authors.affiliation: Carlsberg Laboratory, Department of Chemistry, Valby, Denmark.
authors.forename: K M
authors.initials: KM
authors.lastname: Halkes
authors.forename: C H
authors.initials: CH
authors.lastname: Gotfredsen
authors.forename: M
authors.initials: M
authors.lastname: Grøtli
authors.forename: L P
authors.initials: LP
authors.lastname: Miranda
authors.forename: J O
authors.initials: JO
authors.lastname: Duus
authors.forename: M
authors.initials: M
authors.lastname: Meldal
mesh_terms: D000595:Amino Acid Sequence; D002236:Carbohydrate Conformation; D006020:Glycopeptides / Q000138:chemical synthesis* / Q000737:chemistry*; D006031:Glycosylation; D007202:Indicators and Reagents; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D010455:Peptides / Q000737:chemistry*; D011487:Protein Conformation; D012694:Serine; D013912:Threonine; D014443:Tyrosine
chemical_list: D006020:Glycopeptides; D007202:Indicators and Reagents; D010455:Peptides; D013912:Threonine; D014443:Tyrosine; D012694:Serine