Konno Sho, Ishikawa Fumihiro, Suzuki Takehiro, Dohmae Naoshi, Kakeya Hideaki, Tanabe Genzoh
Department of System Chemotherapy and Molecular Sciences, Division of Bioinformatics and Chemical Genomics, Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo, Kyoto, 606-8501, Japan.
Present address: Faculty of Pharmacy, Kindai University, 3-4-1 Kowakae, Higashi-Osaka, Osaka, 577-8502, Japan.
Chembiochem. 2017 Sep 19;18(18):1855-1862. doi: 10.1002/cbic.201700301. Epub 2017 Aug 17.
Phosphopantetheinylation is an essential post-translational protein modification to primary and secondary metabolic pathways that ensures bacterial cell viability and virulence, and it is used in the production of many pharmaceuticals. Traditional methods have not provided a comprehensive understanding of these modifications. By using chemical proteomic probes for adenylation and thiolation domains in nonribosomal peptide synthetases (NRPSs), chemoproteomics has been applied to survey and validate the cellular activity of 4-[3-chloro-5-(trifluoromethyl)pyridin-2-yl]-N-(4-methoxypyridin-2-yl)piperazine-1-carbothioamide (ML267), which is a potent and selective small-molecule 4'-phosphopantetheinyl transferase (PPTase) inhibitor that attenuates secondary metabolism and viability of bacterial cells. ML267 inhibited Sfp-type PPTase and antagonized phosphopantetheinylation in cells, which resulted in a decrease in phosphopantetheinylated NRPSs and the attenuation of Sfp-PPTase-dependent metabolite production. These results indicate that this chemoproteomics platform should enable a precise interpretation of the cellular activities of Sfp-type PPTase inhibitors.
磷酸泛酰巯基乙胺化是对初级和次级代谢途径至关重要的一种翻译后蛋白质修饰，它确保细菌细胞的活力和毒力，并且被用于多种药物的生产。传统方法尚未对这些修饰有全面的了解。通过使用针对非核糖体肽合成酶（NRPSs）中腺苷化和硫醇化结构域的化学蛋白质组学探针，化学蛋白质组学已被应用于研究和验证4-[3-氯-5-(三氟甲基)吡啶-2-基]-N-(4-甲氧基吡啶-2-基)哌嗪-1-碳硫酰胺（ML267）的细胞活性，ML267是一种强效且具有选择性的小分子4'-磷酸泛酰巯基乙胺基转移酶（PPTase）抑制剂，可减弱细菌细胞的次级代谢和活力。ML267抑制Sfp型PPTase并拮抗细胞中的磷酸泛酰巯基乙胺化，这导致磷酸泛酰巯基乙胺化的NRPSs减少以及Sfp-PPTase依赖性代谢产物生成的减弱。这些结果表明，这个化学蛋白质组学平台应该能够精确诠释Sfp型PPTase抑制剂的细胞活性。
