Chemoenzymatic Late-Stage Modifications Enable Downstream Click-Mediated Fluorescent Tagging of Peptides
Chemoenzymatic Late-Stage Modifications Enable Downstream Click-Mediated Fluorescent Tagging of Peptides
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化学酶后期修饰实现下游点击介导的肽荧光标记
DOI：
10.1002/ange.202215979
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发表时间：
2023
影响因子：
--
通讯作者：
Colombano A
中科院分区：
文献类型：
--
作者：
Colombano A
氰基丁酸生物合成途径中的芳香族戊烯基转移酶催化异戊二烯供体的异戊烯基团和香叶基在分子内的化学选择性和区域选择性转移到这些大环和线状多肽中的富电子位置。这些酶通常表现出松弛的底物专一性，被认为是多肽结构多样化的有用生物催化剂。在这里，我们使用22个人工合成的烷基焦磷酸类似物的文库来评估来自Anacycalide A8P途径的N1-色氨酸戊烯基转移酶ACYF的异戊二烯供体特异性，其中许多类似物显示出可进行额外官能化的反应基团。我们进一步使用ACYF在含有色氨酸的环肽中引入反应部分，然后使用点击化学方法对酶修饰的环肽进行荧光标记。这种化学酶策略允许对多肽进行后期修饰，并且在许多应用中都很有用。
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.