Synthesis of cholera toxin B subunit glycoconjugates using site-specific orthogonal oxime and sortase ligation reactions.

Synthesis of cholera toxin B subunit glycoconjugates using site-specific orthogonal oxime and sortase ligation reactions.
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DOI:
10.3389/fchem.2022.958272
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发表时间:
2022
影响因子:
5.5
通讯作者:
--
中科院分区:
化学3区
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报道了一系列双N端和C端功能化霍乱毒素B亚单位(CTB)糖结合物的化学酶促合成方法。采用固相法合成具有不同TN抗原糖基化水平的粘蛋白1多肽[MUC1(TN)]。利用索尔特酶介导的连接,MUC1(Tn)表位以明确的方式连接到CTB的C末端,从而允许高密度展示MUC1(Tn)表位。这项工作探索了结合糖肽使用山梨酸酶介导的连接的挑战和获得高水平结合的实际考虑。此外,我们还描述了将两种正交标记方法
The chemoenzymatic synthesis of a series of dual N- and C-terminal–functionalized cholera toxin B subunit (CTB) glycoconjugates is described. Mucin 1 peptides bearing different levels of Tn antigen glycosylation [MUC1(Tn)] were prepared via solid-phase peptide synthesis. Using sortase-mediated ligation, the MUC1(Tn) epitopes were conjugated to the C-terminus of CTB in a well-defined manner allowing for high-density display of the MUC1(Tn) epitopes. This work explores the challenges of using sortase-mediated ligation in combination with glycopeptides and the practical considerations to obtain high levels of conjugation. Furthermore, we describe methods to combine two orthogonal labeling methodologies, oxime- and sortase-mediated ligation, to expand the biochemical toolkit and produce dual N- and C-terminal–labeled conjugates.
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