A novel Gal(β1-4)Gal(β1-4)Fuc(α1-6)-core modification attached to the proximal N-acetylglucosamine of keyhole limpet haemocyanin (KLH) N-glycans
A novel Gal(β1-4)Gal(β1-4)Fuc(α1-6)-core modification attached to the proximal N-acetylglucosamine of keyhole limpet haemocyanin (KLH) N-glycans
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DOI：
10.1042/bj20031380
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发表时间：
2004-03-01
影响因子：
4.1
通讯作者：
Hokke, CH
中科院分区：
文献类型：
--
作者：
Wuhrer, M;Robijn, MLM;Hokke, CH
KLH（锁孔帽贝血青素）是海蜗牛megthura crenulata的载氧分子，常被用作佐剂或半抗原载体，用于多肽、低聚糖或其他低分子质量有机化合物的免疫。KLH表现出几种碳水化合物决定因素，其中至少有一些是免疫原性的：它与血吸虫共享一个抗原的Fuc(α 1-3) GalNAc-决定因素，并且在其n-聚糖上含有独特的Gal(β 1-6)Man-结构基元。该研究揭示了n -聚糖中含有不寻常的+/-Gal(β 1-4)Gal(β 1-4) Fuc-单元(α 1-6)-与n -乙酰氨基葡萄糖残基还原端连接的存在。通过连锁分析、外糖苷酶酶切、矩阵辅助激光解吸电离-串联质谱和纳米LC-ESI-IT-质谱（LC代表液相色谱，ESI代表电喷雾电离，IT代表离子阱）推断出以下KLH n-聚糖的新结构：Man(α 1-6) [+/- Man(α 1-3)]Man(β 1-4)GlcNAc(β 1-4)[Gal(β 1-4)Fuc(α 1-6)]GlcNAc和Man(α 1-6)Man (β 1-4)GlcNAc(β 1-4)[Gal(1-4)Gal(p 1-4)Fuc(α 1-6)]GlcNAc。Gal(β 1-4)Fuc-和Gal(β 1-4)Gal(β 1-4)Fuc-核心修饰预计具有免疫原性，类似于其他非哺乳动物型核心修饰，并有助于KLH的免疫刺激特性。
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(alpha 1-3) GalNAc- determinant with schistosomes and contains unique Gal( beta 1-6)Man- structural motifs on its N-glycans. This study reveals the presence of N-glycans with unusual +/-Gal(beta 1-4)Gal(beta 1-4) Fuc- units (alpha 1-6)-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(alpha 1-6) [+/- Man (alpha 1-3)]Man(beta 1-4)GlcNAc(beta 1-4)[Gal(beta 1-4)Fuc(alpha 1-6)]GlcNAc and Man(alpha 1-6)Man (beta 1-4)GlcNAc(beta 1-4) [Gal ( 1-4)Gal(p 1-4) Fuc(alpha 1-6)]GlcNAc. The Gal(beta 1-4)Fuc- and Gal(beta 1-4)Gal(beta 1-4)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.