Oligosaccharide microarrays for high-throughput detection and specificity assignments of carbohydrate-protein interactions
Oligosaccharide microarrays for high-throughput detection and specificity assignments of carbohydrate-protein interactions
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DOI：
10.1038/nbt735
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发表时间：
2002-10-01
影响因子：
46.9
通讯作者：
Chai, WG
中科院分区：
文献类型：
--
作者：
Fukui, S;Feizi, T;Chai, WG
我们将寡糖微阵列描述为新糖脂及其在硝化纤维素上的稳健展示。该阵列源自糖蛋白、糖脂、蛋白聚糖、多糖、整个器官或化学合成的寡糖。我们表明，碳水化合物识别蛋白不仅在同质寡糖阵列中而且在异质寡糖阵列中挑选出其配体。初步应用揭示了新的发现，包括：（i）在大脑中的O-聚糖中，抗L5识别的基于N-乙酰基乳糖胺的Lewis x序列相对丰富，而抗SSEA-1识别的基于聚-N-乙酰基乳糖胺的Lewis x序列较少； (ii) 对硫酸软骨素抗血清和抗体 (CS-56) 识别的硫酸软骨素寡糖的深入了解； (iii) 细胞因子干扰素-γ (IFN-γ) 和趋化因子 RANTES 与硫酸化序列（例如 HNK-1、磺基-Lewis(x) 和磺基-Lewis(a)）以及糖胺聚糖的结合。该方法为在蛋白质组中发现新的碳水化合物识别蛋白以及绘制糖组中碳水化合物识别结构的全部内容开辟了道路。
We describe microarrays of oligosaccharides as neoglycolipids and their robust display on nitrocellulose. The arrays are sourced from glycoproteins, glycolipids, proteoglycans, polysaccharides, whole organs, or from chemically synthesized oligosaccharides. We show that carbohydrate- recognizing proteins single out their ligands not only in arrays of homogeneous oligosaccharides but also in arrays of heterogeneous oligosaccharides. Initial applications have revealed new findings, including: (i) among O-glycans in brain, a relative abundance of the Lewis x sequence based on N-acetyllactosamine recognized by anti-L5, and a paucity of the Lewis x sequence based on poly-N-acetyllactosamine recognized by anti-SSEA-1; (ii) insights into chondroitin sulfate oligosaccharides recognized by an antiserum and an antibody (CS-56) to chondroitin sulfates; and (iii) binding of the cytokine interferon-gamma (IFN-gamma) and the chemokine RANTES to sulfated sequences such as HNK-1, sulfo-Lewis(x), and sulfo-Lewis(a), in addition to glycosaminoglycans. The approach opens the way for discovering new carbohydrate- recognizing proteins in the proteome and for mapping the repertoire of carbohydrate recognition structures in the glycome.