Data_Sheet_1_A Comprehensive Analysis of the Lysine Acetylome in the Aquatic Animals Pathogenic Bacterium Vibrio mimicus.docx

Protein lysine acetylation is an evolutionarily conserved post-translational modification (PTM), which is dynamic and reversible, playing a crucial regulatory role in almost every aspect of metabolism, of both eukaryotes and prokaryotes. Several global lysine acetylome studies have been carried out in various bacteria, but thus far, there have been no reports of lysine acetylation for the commercially important aquatic animal pathogen Vibrio mimicus. In the present study, we used anti-Ac-K antibody beads to highly sensitive immune-affinity purification and combined high-resolution LC-MS/MS to perform the first global lysine acetylome analysis in V. mimicus, leading to the identification of 1,097 lysine-acetylated sites on 582 proteins, and more than half (58.4%) of the acetylated proteins had only one site. The analysis of acetylated modified peptide motifs revealed six significantly enriched motifs, namely, KacL, KacR, L(-2) KacL, LKacK, L(-7) EKac, and IEKac. In addition, bioinformatic assessments state clearly that acetylated proteins have a hand in many important biological processes in V. mimicus, such as purine metabolism, ribosome, pyruvate metabolism, glycolysis/gluconeogenesis, the TCA cycle, and so on. Moreover, 13 acetylated proteins were related to the virulence of V. mimicus. To sum up, this is a comprehensive analysis whole situation protein lysine acetylome in V. mimicus and provides an important foundation for in-depth study of the biological function of lysine acetylation in V. mimicus.

蛋白质赖氨酸乙酰化（Protein lysine acetylation）是一种进化上保守的翻译后修饰（post-translational modification, PTM），具有动态可逆的特性，在真核生物与原核生物的几乎所有代谢过程中均发挥关键调控作用。目前已有多项针对多种细菌的全局赖氨酸乙酰化组研究，但迄今尚无关于具有重要商业价值的水生动物病原菌拟态弧菌（Vibrio mimicus）赖氨酸乙酰化的相关报道。本研究采用抗Ac-K抗体磁珠进行高灵敏度免疫亲和纯化，并结合高分辨率液相色谱-串联质谱（LC-MS/MS），首次对拟态弧菌的全局赖氨酸乙酰化组进行分析，最终在582个蛋白质上鉴定到1097个赖氨酸乙酰化位点，其中超过半数（58.4%）的乙酰化蛋白质仅含1个乙酰化位点。对乙酰化修饰肽基序的分析共鉴定出6种显著富集的基序，分别为KacL、KacR、L(-2)KacL、LKacK、L(-7)EKac及IEKac。此外，生物信息学分析明确显示，乙酰化蛋白质参与拟态弧菌的诸多重要生物学过程，包括嘌呤代谢、核糖体、丙酮酸代谢、糖酵解/糖异生以及三羧酸循环（TCA cycle）等。另有13个乙酰化蛋白质与拟态弧菌的毒力相关。综上，本研究全面解析了拟态弧菌的蛋白质赖氨酸乙酰化组全貌，为深入研究拟态弧菌中赖氨酸乙酰化的生物学功能提供了重要基础。