Reductive site-selective atypical C,Z-type/N2-C2 cleavage allows C-terminal protein amidation.
Reductive site-selective atypical C,Z-type/N2-C2 cleavage allows C-terminal protein amidation.
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DOI：
10.1126/sciadv.abl8675
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发表时间：
2022-04-08
期刊：
影响因子：
13.6
通讯作者：
Davis BG
中科院分区：
文献类型：
--
作者：
Mollner TA;Giltrap AM;Zeng Y;Demyanenko Y;Buchanan C;Oehlrich D;Baldwin AJ;Anthony DC;Mohammed S;Davis BG
生物分子环境可以通过介导和调节自我修饰（例如，自裂解）的潜力来增强化学反应。虽然在某些生物分子（例如RNA核酶）中发现了这些增强模式，但在蛋白质中更为罕见。靶向蛋白水解裂解对生理学、生物技术甚至新兴疗法都是至关重要的。然而，纯化学诱导的蛋白质位点选择性切割方法仍然很少。在这里，作为原理证明，我们设计并测试了一个系统，旨在将蛋白质增强化学与标签修饰相结合，以实现由二硼促进的合成还原性蛋白质化学。这种还原驱动的单电子化学现在可以在水条件下和细胞裂解物中实现操作简单，位点选择性的切割方案，用于蛋白质直接作为易于获得的脱氢丙氨酸（Dha）残基作为标签。通过这种方式，一种温和、高效、无酶的方法现在不仅可以进行精确的化学蛋白质水解，还可以同时用于去除亲和标签和/或蛋白质末端编辑，以创建改变的N和c末端，如蛋白质酰胺化（─CONH2）。一种仿生还原试剂在非肽键上切割蛋白质，留下可以改变功能的酰胺端。
Biomolecule environments can enhance chemistries with the potential to mediate and modulate self-modification (e.g., self-cleavage). While these enhanced modes are found in certain biomolecules (e.g., RNA ribozymes), it is more rare in proteins. Targeted proteolytic cleavage is vital to physiology, biotechnology, and even emerging therapy. Yet, purely chemically induced methods for the site-selective cleavage of proteins remain scarce. Here, as a proof of principle, we designed and tested a system intended to combine protein-enhanced chemistry with tag modification to enable synthetic reductive protein chemistries promoted by diboron. This reductively driven, single-electron chemistry now enables an operationally simple, site-selective cleavage protocol for proteins directed to readily accessible dehydroalanine (Dha) residues as tags under aqueous conditions and in cell lysates. In this way, a mild, efficient, enzyme-free method now allows not only precise chemical proteolysis but also simultaneous use in the removal of affinity tags and/or protein-terminus editing to create altered N- and C-termini such as protein amidation (─CONH2). A biomimetic reductive reagent cleaves proteins at a nonpeptidic bond to leave amide termini that can switch function.