Fujian Provincial Key Laboratory of Agroecological Processing and Safety Monitoring (School of Life Sciences, Fujian Agriculture and Forestry University), Fuzhou, PR China;; Key Laboratory of Crop Ecology and Molecular Physiology (Fujian Agriculture and Forestry University), Fujian Province University, Fuzhou, PR China;; Shanghai Key Laboratory of Plant Functional Genomics and Resources, Shanghai Chenshan Plant Science Research Center, Chinese Academy of Sciences, Shanghai Chenshan Botanical Garden, Shanghai, PR China.
Fujian Provincial Key Laboratory of Agroecological Processing and Safety Monitoring (School of Life Sciences, Fujian Agriculture and Forestry University), Fuzhou, PR China;; Key Laboratory of Crop Ecology and Molecular Physiology (Fujian Agriculture and Forestry University), Fujian Province University, Fuzhou, PR China.
Mol Cell Proteomics. 2019 Feb;18(2):200-215. doi: 10.1074/mcp.RA118.001035. Epub 2018 Oct 23.
Protein modification by lysine succinylation is a newly identified post-translational modification (PTM) of lysine residues and plays an important role in diverse physiological functions, although their associated biological characteristics are still largely unknown. Here, we investigated the effects of lysine succinylation on the physiological regulation within a well-known fish pathogen, A high affinity purification method was used to enrich peptides with lysine succinylation in ATCC 7966, and a total of 2,174 lysine succinylation sites were identified on 666 proteins using LC-MS/MS. Gene ontology analysis indicated that these succinylated proteins are involved in diverse metabolic pathways and biological processes, including translation, protein export, and central metabolic pathways. The modifications of several selected candidates were further validated by Western blotting. Using site-directed mutagenesis, we observed that the succinylation of lysines on S-ribosylhomocysteine lyase (LuxS) at the K23 and K30 sites positively regulate the production of the quorum sensing autoinducer AI-2, and that these PTMs ultimately alter its competitiveness with another pathogen, Moreover, subsequent metabolomic analyses indicated that K30 succinylation on LuxS may suppress the activated methyl cycle (AMC) and that both the K23 and K30 sites are involved in amino acid metabolism. Taken together, the results from this study provide significant insights into the functions of lysine succinylation and its critical roles on LuxS in regulating the cellular physiology of
赖氨酸琥珀酰化修饰是一种新发现的赖氨酸残基翻译后修饰（PTM），在多种生理功能中发挥着重要作用，但其相关生物学特性尚不清楚。在这里，我们研究了赖氨酸琥珀酰化对一种著名鱼类病原体生理调节的影响。采用高亲和力的纯化方法从 ATCC 7966 中富集具有赖氨酸琥珀酰化的肽段，通过 LC-MS/MS 共鉴定了 666 种蛋白质上的 2174 个赖氨酸琥珀酰化位点。GO 分析表明，这些琥珀酰化蛋白参与了多种代谢途径和生物学过程，包括翻译、蛋白质输出和中心代谢途径。通过 Western blot 进一步验证了几个候选蛋白的修饰。通过定点突变，我们观察到 S-核糖基同型半胱氨酸裂解酶（LuxS）上赖氨酸的琥珀酰化修饰（K23 和 K30）可正向调节群体感应自动诱导物 AI-2 的产生，这些 PTM 最终改变了其与另一种病原体的竞争力。此外，随后的代谢组学分析表明，LuxS 上的 K30 琥珀酰化可能抑制激活甲基循环（AMC），并且 K23 和 K30 位点都参与了氨基酸代谢。总之，本研究结果为赖氨酸琥珀酰化及其在调节 LuxS 细胞生理功能中的关键作用提供了重要的见解。
