Structural basis for multiple sugar recognition of Jacalin-related human ZG16p lectin.
Structural basis for multiple sugar recognition of Jacalin-related human ZG16p lectin.
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DOI：
10.1074/jbc.m113.539114
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发表时间：
2014-06-13
期刊：
影响因子：
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通讯作者：
Yamaguchi Y
中科院分区：
文献类型：
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作者：
Kanagawa M;Liu Y;Hanashima S;Ikeda A;Chai W;Nakano Y;Kojima-Aikawa K;Feizi T;Yamaguchi Y
背景：ZG 16 p是一种可溶性哺乳动物凝集素，具有Jacalin相关的β-prism折叠。结果如下：ZG 16 p分别通过典型的浅甘露糖结合口袋和相邻的碱性表面积与短α-甘露糖相关聚糖和糖胺聚糖结合。结论：ZG 16 p具有独特的多配体结合特性。重要性：ZG 16 p配体结合的结构见解对于理解蛋白质的生物学功能至关重要。ZG 16 p是一种可溶性哺乳动物凝集素，首次被描述为具有Jacalin相关的β-prism折叠。据报道，ZG 16 p结合糖胺聚糖和甘露糖。为了确定多种糖结合特性的结构基础，我们对人ZG 16 p进行了聚糖微阵列分析。我们观察到ZG 16 p优先结合α-甘露糖终止的短聚糖，如Ser/Thr连接的O-甘露糖，但不结合高甘露糖型N-聚糖。在检查的硫酸化糖胺聚糖低聚物中，硫酸软骨素B和肝素寡糖显示出显著结合。在选定的配体存在下，人ZG 16 p凝集素的晶体学研究揭示了多糖识别的机制。Manα1- 3 Man和Glcβ1-3Glc以不同的方向结合：前者的非还原端和后者的还原端适合于典型的浅甘露糖结合口袋。使用15 N标记的ZG 16 p的溶液NMR分析定义了肝素结合区域，其位于蛋白质的相邻平坦表面上。阵列上竞争性结合测定表明，ZG 16 p有可能同时结合两种类型的配体。ZG 16 p对广谱配体的识别可能解释了该凝集素在通过糖胺聚糖结合形成酶原颗粒以及通过α-甘露糖相关识别在消化系统中识别病原体中的多种功能。
Background: ZG16p is a soluble mammalian lectin with a Jacalin-related β-prism-fold. Results: ZG16p binds to short α-mannose-related glycans and glycosaminoglycans via the canonical shallow mannose-binding pocket and an adjacent basic surface area, respectively. Conclusion: ZG16p possesses a unique feature of multiple-ligand binding. Significance: Structural insights of ZG16p ligand binding are crucial for understanding the biological functions of the protein. ZG16p is a soluble mammalian lectin, the first to be described with a Jacalin-related β-prism-fold. ZG16p has been reported to bind both to glycosaminoglycans and mannose. To determine the structural basis of the multiple sugar-binding properties, we conducted glycan microarray analyses of human ZG16p. We observed that ZG16p preferentially binds to α-mannose-terminating short glycans such as Ser/Thr-linked O-mannose, but not to high mannose-type N-glycans. Among sulfated glycosaminoglycan oligomers examined, chondroitin sulfate B and heparin oligosaccharides showed significant binding. Crystallographic studies of human ZG16p lectin in the presence of selected ligands revealed the mechanism of multiple sugar recognition. Manα1–3Man and Glcβ1–3Glc bound in different orientations: the nonreducing end of the former and the reducing end of the latter fitted in the canonical shallow mannose binding pocket. Solution NMR analysis using 15N-labeled ZG16p defined the heparin-binding region, which is on an adjacent flat surface of the protein. On-array competitive binding assays suggest that it is possible for ZG16p to bind simultaneously to both types of ligands. Recognition of a broad spectrum of ligands by ZG16p may account for the multiple functions of this lectin in the formation of zymogen granules via glycosaminoglycan binding, and in the recognition of pathogens in the digestive system through α-mannose-related recognition.