School of Pharmaceutical Science and Technology, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences, Hangzhou, China.
State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China.
Methods Mol Biol. 2023;2676:131-146. doi: 10.1007/978-1-0716-3251-2_9.
Posttranslational modifications (PTMs) of lysine residues are major regulators of gene expression, protein-protein interactions, and protein localization and degradation. Histone lysine benzoylation is a recently identified epigenetic marker associated with active transcription, which has physiological relevance distinct from histone acetylation and can be regulated by debenzoylation of sirtuin 2 (SIRT2). Herein, we provide a protocol for the incorporation of benzoyllysine and fluorinated benzoyllysine into full-length histone proteins, which further serve as benzoylated histone probes with NMR or fluorescence signal for investigating the dynamics of SIRT2-mediated debenzoylation.
赖氨酸残基的翻译后修饰（PTMs）是基因表达、蛋白质-蛋白质相互作用以及蛋白质定位和降解的主要调节剂。组蛋白赖氨酸苯甲酰化是最近发现的与转录活性相关的表观遗传标记，它与组蛋白乙酰化具有不同的生理相关性，并且可以通过 SIRT2（沉默信息调节因子 2）的去苯甲酰化来调节。本文提供了一种将苯甲酰化赖氨酸和氟化苯甲酰化赖氨酸掺入全长组蛋白蛋白中的方案，这些组蛋白蛋白进一步作为苯甲酰化组蛋白探针，具有 NMR 或荧光信号，用于研究 SIRT2 介导的去苯甲酰化动力学。
