Synthesis of biologically active N- and O-linked glycans with multisialylated poly-N-acetyllactosamine extensions using P. damsela α2-6 sialyltransferase.
Synthesis of biologically active N- and O-linked glycans with multisialylated poly-N-acetyllactosamine extensions using P. damsela α2-6 sialyltransferase.
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DOI：
10.1021/ja409781c
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发表时间：
2013-12-11
影响因子：
15
通讯作者：
Paulson JC
中科院分区：
文献类型：
--
作者：
Nycholat CM;Peng W;McBride R;Antonopoulos A;de Vries RP;Polonskaya Z;Finn MG;Dell A;Haslam SM;Paulson JC
N-和O-连接糖蛋白上的唾液酸苷在许多生物过程中起着重要作用，合成聚糖探针已被证明是阐明这些功能的有价值的工具。虽然唾液酸通常与末端非还原性半乳糖α2-3或α2-6连接，但据报道，从大鼠唾液腺和人结肠粘蛋白中分离的聚LacNAc延伸的核心-3 O-连接聚糖含有多个内部Neu 5Ac α2- 6 Gal表位。在这里，我们开发了一种有效的方法来合成具有多唾液酸化聚LacNAc延伸的N-和O-连接聚糖的文库，包括天然存在的多唾液酸化核心-3 O-连接聚糖。我们发现来自亮杆菌（Photobacterium damsela）的重组α2-6唾液酸转移酶（Pd 2，6ST）具有独特的区域选择性，并且能够唾液酸化poly-LacNAc延伸聚糖中的内部半乳糖残基，这通过MS/MS分析得到证实。使用展示该文库的聚糖微阵列，我们发现Neu 5Ac α2- 6 Gal特异性流感病毒血凝素、单细胞凝集素和植物凝集素在很大程度上不受相邻内部唾液酸化的影响，并且在一些情况下，内部唾液酸被识别为配体。免疫鸡可获得特异性抗唾液酸苷内表位的多克隆IgY抗体。
Sialosides on N- and O-linked glycoproteins play a fundamental role in many biological processes and synthetic glycan probes have proven to be valuable tools for elucidating these functions. Though sialic acids are typically found α2-3 or α2-6-linked to a terminal non-reducing end galactose, poly-LacNAc extended core-3 O-linked glycans isolated from rat salivary glands and human colonic mucins have been reported to contain multiple internal Neu5Acα2-6Gal epitopes. Here, we have developed an efficient approach for the synthesis of a library of N- and O-linked glycans with multi-sialylated poly-LacNAc extensions, including naturally occurring multi-sialylated core-3 O-linked glycans. We have found that a recombinant α2-6 sialyltransferase from Photobacterium damsela (Pd2,6ST) exhibits unique regioselectivity and is able to sialylate internal galactose residues in poly-LacNAc extended glycans which was confirmed by MS/MS analysis. Using a glycan microarray displaying this library, we found that Neu5Acα2-6Gal specific influenza virus hemagglutinins, siglecs and plant lectins are largely unaffected by adjacent internal sialylation, and in several cases the internal sialic acids are recognized as ligands. Polyclonal IgY antibodies specific for internal sialoside epitopes were elicited in inoculated chickens.
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影响因子：
16.6
作者：
Yu, Hai;Huang, Shengsu;Chen, Xi
通讯作者：
Chen, Xi
影响因子：
3.6
作者：
Blixt, O;Brown, J;Paulson, JC
通讯作者：
Paulson, JC
DOI：
10.1097/aci.0b013e32835b594a
发表时间：
2013-02
影响因子：
2.8
作者：
Kiwamoto T;Katoh T;Tiemeyer M;Bochner BS
通讯作者：
Bochner BS
影响因子：
3.6
作者：
Kajihara, Y;Yamamoto, T;Terada, I
通讯作者：
Terada, I
DOI：
10.1007/s00018-005-5589-y
发表时间：
2006-06
期刊：
Cellular and molecular life sciences : CMLS
影响因子：
--
作者：
Lehmann F;Tiralongo E;Tiralongo J
通讯作者：
Tiralongo J