Synthesis and preliminary evaluation of neoglycopolymers carrying multivalent N-glycopeptide units.
Synthesis and preliminary evaluation of neoglycopolymers carrying multivalent N-glycopeptide units.
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携带多价N-糖肽单元的新糖聚合物的合成和初步评价。
DOI：
10.1016/j.ijbiomac.2019.09.255
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Kimura Y.
中科院分区：
文献类型：
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作者：
Takeda N;Maeda M;Itano S;Takase M;Kimura M;Kimura Y.
在本研究中，为了发现植物中未表征的聚糖结合受体或凝集素样受体，我们开发了三种类型的N-糖肽缀合的neoglycopolymers;第一种与植物复合型N-聚糖（M3 FX），第二种与高甘露糖型N-聚糖（M8），第三种与动物复合型N-聚糖（NeuAc 2Gal 2GN 2 M3）。以银杏、豇豆种子贮藏糖蛋白为原料，用放线菌素酶解卵黄糖肽，制备了三种不同的天冬酰胺寡糖（Asn-M3 FX、Asn-M8、Asn-NeuAc 2Gal 2GN 2 M3）。以4-（4，6-二甲氧基-1，3，5-三嗪-2-基）-4-甲基吗啉盐酸盐（DMT-MM）为偶联剂，将天冬酰胺寡糖的α-氨基与聚γ-L-谷氨酸（γ-L-PGA）的羧基偶联，合成了新糖共聚物。通过凝胶过滤和反相HPLC的组合纯化所得的neoglycopolymers。通过酸水解后的氨基酸组成分析来估计N-聚糖掺入γ-L-PGA（mol%）。Asn-M3 FX、Asn-M8和Asn-NeuAc 2Gal 2GN 2 M3与γ-L-PGA的结合率分别为15.4%、8.6%和11.1%，表明分别有近890、500和640个N-聚糖分子与γ-L-PGA结合。此外，我们证实，neoglycopolymer携带的多价高甘露糖型N-聚糖是一个有用的工具，从刀豆提取物中的甘露糖结合蛋白，伴刀豆球蛋白A的快速纯化。
In the present study, for the discovery of uncharacterized glycan-binding receptors or lectin-like receptors in plants, we developed neoglycopolymers to which three types ofN-glycopeptides are conjugated; the first with plant complex typeN-glycan (M3FX), the second with high-mannose typeN-glycan (M8), and the third with animal complex typeN-glycan (NeuAc2Gal2GN2M3). Three types of Asn-oligosaccharide (Asn-M3FX, Asn-M8, or Asn-NeuAc2Gal2GN2M3) were prepared from storage glycoproteins ofGinkgo bilobaseeds,Vignaangularisseeds, and egg yolk glycopeptides from actinase digests of each glycoproteins or glycopeptide. Neoglycopolymers were synthesized such that the α-amino groups of Asn-oligosaccharide were coupled to the carboxyl groups of poly-γ-L-glutamic acid (γ-L-PGA) with 4-(4,6-Dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloriden-hydrate (DMT-MM). The resulting neoglycopolymers were purified through a combination of gel-filtration and reverse-phase HPLC. The incorporation ofN-glycans into γ-L-PGA (mol%) was estimated through amino acid composition analysis after acid hydrolysis. The incorporation rates of Asn-M3FX, Asn-M8, and Asn-NeuAc2Gal2GN2M3 into γ-L-PGA were 15.4%, 8.6%, and 11.1%, indicating that nearly 890, 500, and 640 molecules ofN-glycans were conjugated with γ-L-PGA, respectively. Furthermore, we confirmed that the neoglycopolymer carrying the multivalent high-mannose typeN-glycans is a useful tool for rapid purification of mannose-binding protein, Concanavalin A, from jack bean extract.