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. 2004 Mar 1;378(Pt 2):625–632. doi: 10.1042/BJ20031380

A novel Gal(beta1-4)Gal(beta1-4)Fuc(alpha1-6)-core modification attached to the proximal N-acetylglucosamine of keyhole limpet haemocyanin (KLH) N-glycans.

Manfred Wuhrer 1, Marjolein L M Robijn 1, Carolien A M Koeleman 1, Crina I A Balog 1, Rudolf Geyer 1, André M Deelder 1, Cornelis H Hokke 1
PMCID: PMC1223963  PMID: 14613482

Abstract

KLH (keyhole limpet haemocyanin), the oxygen-carrying molecule of the marine snail Megathura crenulata, is often used as an adjuvant or as a hapten carrier for immunizations with peptides, oligosaccharides or other low-molecular-mass organic compounds. KLH exhibits several carbohydrate determinants, at least some of which are immunogenic: it shares an antigenic Fuc(alpha1-3)GalNAc-determinant with schistosomes and contains unique Gal-(beta1-6)Man-structural motifs on its N-glycans. This study reveals the presence of N-glycans with unusual +/-Gal(beta1-4)Gal(beta1-4)Fuc- units (alpha1-6)-linked to the reducing end N -acetylglucosamine residue. The following novel structures of KLH N-glycans were deduced by linkage analysis, exoglycosidase digestion, matrix-assisted laser-desorption ionization-tandem MS and nano-LC-ESI-IT-MS (where LC stands for liquid chromatography, ESI for electrospray ionization and IT for ion trap): Man(alpha1-6)[+/-Man(alpha1-3)]Man(beta1-4)GlcNAc(beta1-4)[Gal(beta1-4)Fuc(alpha1-6)]GlcNAc and Man(alpha1-6)Man(beta1-4)GlcNAc(beta1-4)[Gal(beta1-4)Gal(beta1-4)Fuc(alpha1-6)]GlcNAc. The Gal(beta1-4)Fuc- and Gal(beta1-4)Gal(beta1-4)Fuc- core modifications are expected to be immunogenic, similar to other non-mammalian-type core modifications, and to contribute to the immunostimulatory properties of KLH.

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Selected References

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