Superbacteria Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea.
Proteomics. 2013 May;13(10-11):1726-36. doi: 10.1002/pmic.201200001.
N(ε) -lysine acetylation, a reversible and highly regulated PTM, has been shown to occur in the model Gram-negative bacteria Escherichia coli and Salmonella enterica. Here, we extend this acetylproteome analysis to Bacillus subtilis, a model Gram-positive bacterium. Through anti-acetyllysine antibody-based immunoseparation of acetylpeptides followed by nano-HPLC/MS/MS analysis, we identified 332 unique lysine-acetylated sites on 185 proteins. These proteins are mainly involved in cellular housekeeping functions such as central metabolism and protein synthesis. Fifity-nine of the lysine-acetylated proteins showed homology with lysine-acetylated proteins previously identified in E. coli, suggesting that acetylated proteins are more conserved. Notably, acetylation was found at or near the active sites predicted by Prosite signature, including SdhA, RocA, Kbl, YwjH, and YfmT, indicating that lysine acetylation may affect their activities. In 2-amino-3-ketobutyrate CoA ligase Kbl, a class II aminotransferase, a lysine residue involved in pyridoxal phosphate attachment was found to be acetylated. This data set provides evidence for the generality of lysine acetylation in eubacteria and opens opportunities to explore the consequences of acetylation modification on the molecular physiology of B. subtilis.
N(ε)-赖氨酸乙酰化是一种可逆且高度调控的翻译后修饰（PTM），已在模式革兰氏阴性菌大肠杆菌和沙门氏菌中得到证实。在这里，我们将这种乙酰化蛋白质组分析扩展到模式革兰氏阳性菌枯草芽孢杆菌。通过基于抗乙酰赖氨酸抗体的乙酰肽免疫分离，然后进行纳升 HPLC/MS/MS 分析，我们在 185 种蛋白质上鉴定出了 332 个独特的赖氨酸乙酰化位点。这些蛋白质主要参与细胞的管家功能，如中心代谢和蛋白质合成。59 种赖氨酸乙酰化蛋白与先前在大肠杆菌中鉴定的赖氨酸乙酰化蛋白具有同源性，表明乙酰化蛋白更为保守。值得注意的是，在 Prosite 特征预测的活性位点或其附近发现了乙酰化，包括 SdhA、RocA、Kbl、YwjH 和 YfmT，表明赖氨酸乙酰化可能影响它们的活性。在 2-氨基-3-酮丁酸辅酶 A 连接酶 Kbl（一种 II 类氨基转移酶）中，与吡哆醛磷酸酯结合有关的一个赖氨酸残基被发现发生了乙酰化。该数据集为真细菌中赖氨酸乙酰化的普遍性提供了证据，并为探索乙酰化修饰对枯草芽孢杆菌分子生理学的影响提供了机会。
