Solid-phase glycosylation of peptide templates and on-bead MAS-NMR analysis: perspectives for glycopeptide libraries.
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.
Halkes
K M
KM
Carlsberg Laboratory, Department of Chemistry, Valby, Denmark.
Gotfredsen
C H
CH
Grøtli
M
M
Miranda
L P
LP
Duus
J O
JO
Meldal
M
M
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
D006020:Glycopeptides; D007202:Indicators and Reagents; D010455:Peptides; D013912:Threonine; D014443:Tyrosine; D012694:Serine