Chemoenzymatic Late-Stage Modifications Enable Downstream Click-Mediated Fluorescent Tagging of Peptides.
Chemoenzymatic Late-Stage Modifications Enable Downstream Click-Mediated Fluorescent Tagging of Peptides.
复制标题
化学酶后期修饰可实现下游点击介导的肽荧光标记。
DOI：
10.1002/anie.202215979
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Colombano A
中科院分区：
文献类型：
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作者：
Colombano A
来自氰菌素生物合成途径的芳香族异戊二烯基转移酶催化异戊二烯供体的异戊二烯基团的化学选择性和区域选择性分子内转移到这些大环和线性肽中的富含电子的位置。这些酶通常表现出宽松的底物特异性，并被认为是肽结构多样化的有用生物催化剂。在此，我们使用 22 种合成的烷基焦磷酸类似物库评估了来自阿那环酰胺 A8P 途径的 N1-色氨酸异戊二烯基转移酶 AcyF 的异戊二烯供体特异性，其中许多类似物显示出适合额外功能化的反应基团。我们进一步使用 AcyF 将反应部分引入含色氨酸的环肽中，随后使用点击化学来荧光标记酶促修饰的肽。这种化学酶策略允许对肽进行后期修饰，并且可用于许多应用。
Aromatic prenyltransferases from cyanobactin biosynthetic pathways catalyse the chemoselective and regioselective intramolecular transfer of prenyl/geranyl groups from isoprene donors to an electron‐rich position in these macrocyclic and linear peptides. These enzymes often demonstrate relaxed substrate specificity and are considered useful biocatalysts for structural diversification of peptides. Herein, we assess the isoprene donor specificity of the N1‐tryptophan prenyltransferase AcyF from the anacyclamide A8P pathway using a library of 22 synthetic alkyl pyrophosphate analogues, of which many display reactive groups that are amenable to additional functionalization. We further used AcyF to introduce a reactive moiety into a tryptophan‐containing cyclic peptide and subsequently used click chemistry to fluorescently label the enzymatically modified peptide. This chemoenzymatic strategy allows late‐stage modification of peptides and is useful for many applications.