An oligosaccharide chain attached to the asparagine residue in a structurally defined peptide was produced by an in vitro oligosaccharide- transfer reaction, using membrane fractions that contained the oligosaccharyltransferase from the hyperthermophilic archaeon, Pyrococcus furiosus. The chemical structure of the N-glycan was elucidated by sugar analysis, NMR spectroscopy, and MS spectrometry, which revealed the structure. The shorter glycan structures lacking one or two xylose residues were also transferred by the P. furiosus oligosaccharyltransferase. The archaeal N-glycans are known to exhibit a high degree of structural variation. The structure of the P. furiosus N-glycan is novel and unique. The present data will be useful for structural and functional studies of the P. furiosus oligosaccharyltransferase.
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