瑞士苏黎世联邦理工学院Lang, Kathrin研究团队报道了，利用遗传密码扩展解读蛋白质琥珀酰化和戊二酰化的功能作用。相关研究成果发表在2024年3月26日出版的国际知名学术期刊《自然—化学》。

翻译后修饰（PTMs）动态调节细胞过程。赖氨酸经过一系列酰化反应，包括丙二酰化、琥珀酰化（SucK）和戊二酰化（GluK）。这些PTMs过程增加了赖氨酸侧链的大小，并在生理条件下将其电荷从+1逆转为1，可能会影响蛋白质的结构和功能。

为了理解这些PTMs的功能作用，生化研究需要均匀修饰的蛋白质。虽然通过遗传密码扩展对PTMs及其模拟物的位点特异性编码，促进了许多PTMs功能作用的表征，但带负电荷的赖氨酸酰化却挑战了这种方法。

该文中，研究人员描述了SucK和GluK通过硫酯衍生物，暂时掩盖其负电荷而位点特异性地结合到蛋白质中。研究人员制备了琥珀酰化和戊二酰化的细菌和哺乳动物靶蛋白，包括不可重折叠的多结构域蛋白。这使研究人员能够研究琥珀酰化和戊二酰化如何影响代谢酶的酶活性，并调节从复制到泛素信号传导的生物过程中的蛋白质-DNA和蛋白质-蛋白质相互作用。

附：英文原文

Title: Deciphering functional roles of protein succinylation and glutarylation using genetic code expansion

Author: Weyh, Maria, Jokisch, Marie-Lena, Nguyen, Tuan-Anh, Fottner, Maximilian, Lang, Kathrin

Issue&Volume: 2024-03-26

Abstract: Post-translational modifications (PTMs) dynamically regulate cellular processes. Lysine undergoes a range of acylations, including malonylation, succinylation (SucK) and glutarylation (GluK). These PTMs increase the size of the lysine side chain and reverse its charge from +1 to 1 under physiological conditions, probably impacting protein structure and function. To understand the functional roles of these PTMs, homogeneously modified proteins are required for biochemical studies. While the site-specific encoding of PTMs and their mimics via genetic code expansion has facilitated the characterization of the functional roles of many PTMs, negatively charged lysine acylations have defied this approach. Here we describe site-specific incorporation of SucK and GluK into proteins via temporarily masking their negative charge through thioester derivatives. We prepare succinylated and glutarylated bacterial and mammalian target proteins, including non-refoldable multidomain proteins. This allows us to study how succinylation and glutarylation impact enzymatic activity of metabolic enzymes and regulate protein–DNA and protein–protein interactions in biological processes from replication to ubiquitin signalling.

DOI: 10.1038/s41557-024-01500-5

Source: https://www.nature.com/articles/s41557-024-01500-5