Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences, Shanghai, China; CAS Key Laboratory of Quantitative Engineering Biology, Guangdong Provincial Key Laboratory of Synthetic Genomics and Shenzhen Key Laboratory of Synthetic Genomics, Shenzhen Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China; Key Laboratory of Synthetic Biology, University of Chinese Academy of Sciences, Beijing, China.
Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences, Shanghai, China.
Mol Cell Proteomics. 2021;20:100148. doi: 10.1016/j.mcpro.2021.100148. Epub 2021 Sep 14.
Lysine acylations are reversible and ubiquitous post-translational modifications that play critical roles in regulating multiple cellular processes. In the current study, highly abundant and dynamic acetylation, besides succinylation, was uncovered in a soil bacterium, Streptomyces coelicolor. By affinity enrichment using anti-acetyl-lysine antibody and the following LC-MS/MS analysis, a total of 1298 acetylation sites among 601 proteins were identified. Bioinformatics analyses suggested that these acetylated proteins have diverse subcellular localization and were enriched in a wide range of biological functions. Specifically, a majority of the acetylated proteins were also succinylated in the tricarboxylic acid cycle and protein translation pathways, and the bimodification occurred at the same sites in some proteins. The acetylation and succinylation sites were quantified by knocking out either the deacetylase ScCobB1 or the desuccinylase ScCobB2, demonstrating a possible competitive relationship between the two acylations. Moreover, in vitro experiments using synthetically modified peptides confirmed the regulatory crosstalk between the two sirtuins, which may be involved in the collaborative regulation of cell physiology. Collectively, these results provided global insights into the S. coelicolor acylomes and laid a foundation for characterizing the regulatory roles of the crosstalk between lysine acetylation and succinylation in the future.
赖氨酸酰化是一种可逆且普遍存在的翻译后修饰，在调节多种细胞过程中起着关键作用。在本研究中，除琥珀酰化外，土壤细菌链霉菌中还发现了高度丰富和动态的乙酰化。通过使用抗乙酰化赖氨酸抗体进行亲和富集和随后的 LC-MS/MS 分析，在 601 种蛋白质中鉴定出了 1298 个乙酰化位点。生物信息学分析表明，这些乙酰化蛋白质具有不同的亚细胞定位，并富集在广泛的生物学功能中。具体来说，大多数乙酰化蛋白质在三羧酸循环和蛋白质翻译途径中也被琥珀酰化，并且在一些蛋白质中，双修饰发生在相同的位点。通过敲除去乙酰化酶 ScCobB1 或去琥珀酰化酶 ScCobB2 来定量测定乙酰化和琥珀酰化位点，证明了这两种酰化之间可能存在竞争关系。此外，使用合成修饰肽的体外实验证实了两种 sirtuins 之间的调节串扰，这可能参与了细胞生理学的协同调节。总之，这些结果提供了链霉菌酰基组的全局见解，并为未来表征赖氨酸乙酰化和琥珀酰化之间串扰的调节作用奠定了基础。
