A PEGylated photocleavable auxiliary mediates the sequential enzymatic glycosylation and native chemical ligation of peptides.
A PEGylated photocleavable auxiliary mediates the sequential enzymatic glycosylation and native chemical ligation of peptides.
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DOI：
10.1002/anie.201501517
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发表时间：
2015-06-22
期刊：
影响因子：
--
通讯作者：
Becker CF
中科院分区：
文献类型：
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作者：
Bello C;Wang S;Meng L;Moremen KW;Becker CF
旨在理解糖基化模式在蛋白质功能中的特定作用的研究将大大受益于允许直接获得同质糖蛋白的其他方法。在这里，开发和应用的一种有效的方法，用于合成复杂的同质糖基化的肽的基础上的多功能的光裂解助剂进行了描述。助剂中PEG聚合物的存在使得能够以优异的产率进行连续的酶促糖基化和直接分离。经糖基化修饰的肽可直接用于与肽硫酯的天然化学连接，所述肽硫酯容易通过相应糖基化肽基树脂的直接肼解和随后的氧化获得。连接的糖肽可以通过UV照射顺利地脱保护。在这里，我们将这种方法应用于制备携带一种或多种Tn、T或唾液酸-T抗原的上皮肿瘤标志物MUC 1的变体。
Research aimed at understanding the specific role of glycosylation patterns in protein function would greatly benefit from additional approaches allowing direct access to homogeneous glycoproteins. Here the development and application of an efficient approach for the synthesis of complex homogenously glycosylated peptides based on a multifunctional photocleavable auxiliary is described. The presence of a PEG polymer within the auxiliary enables sequential enzymatic glycosylation and straightforward isolation in excellent yields. The auxiliary-modified peptides can be directly used in native chemical ligations with peptide thioesters easily obtained via direct hydrazinolysis of the respective glycosylated peptidyl resins and subsequent oxidation. The ligated glycopeptides can be smoothly deprotected via UV irradiation. Here we apply this approach to the preparation of variants of the epithelial tumor marker MUC1 carrying one or more Tn, T or sialyl-T antigens.