Groundwater Paul W, Narlawar Rajeshwar, Liao Vivian Wan Yu, Bhattacharya Anusri, Srivastava Shalini, Kunal Kishore, Doddareddy Munikumar, Oza Pratik M, Mamidi Ramesh, Marrs Emma C L, Perry John D, Hibbs David E, Panda Dulal
Faculty of Pharmacy, The University of Sydney , Pharmacy and Bank Building, Science Road, Sydney, NSW 2006, Australia.
Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay , Mumbai 400076, India.
Biochemistry. 2017 Jan 24;56(3):514-524. doi: 10.1021/acs.biochem.6b00879. Epub 2017 Jan 11.
Inhibition of FtsZ assembly has been found to stall bacterial cell division. Here, we report the identification of a potent carbocyclic curcumin analogue (2d) that inhibits Bacillus subtilis 168 cell proliferation by targeting the assembly of FtsZ. 2d also showed potent inhibitory activity (minimum inhibitory concentrations of 2-4 mg/L) against several clinically important species of Gram-positive bacteria, including methicillin-resistant Staphylococcus aureus. In addition, 2d displayed a significantly reduced inhibitory effect on human cervical cancer cells in comparison to its effect on bacterial cells. Using live cell imaging of GFP-FtsZ by confocal microscopy, 2d was found to rapidly perturb the cytokinetic FtsZ rings in Bacillus subtilis cells. The immunofluorescence imaging of FtsZ also showed that 2d destroyed the Z-ring in bacteria within 5 min. Prolonged treatment with 2d produced filamentous bacteria, but 2d had no detectable effect either on the nucleoids or on the membrane potential of bacteria. 2d inhibited FtsZ assembly in vitro, whereas it had minimal effects on tubulin assembly. Interestingly, 2d strongly enhanced the GTPase activity of FtsZ and reduced the GTPase activity of tubulin. Furthermore, 2d bound to purified FtsZ with a dissociation constant of 4.0 ± 1.1 μM, and the binding of 2d altered the secondary structures of FtsZ. The results together suggested that the non-natural curcumin analogue 2d possesses powerful antibacterial activity against important pathogenic bacteria, and the evidence indicates that 2d inhibits bacterial proliferation by targeting FtsZ.
已发现抑制FtsZ组装会使细菌细胞分裂停滞。在此，我们报告鉴定出一种有效的碳环姜黄素类似物（2d），它通过靶向FtsZ的组装来抑制枯草芽孢杆菌168细胞增殖。2d对包括耐甲氧西林金黄色葡萄球菌在内的几种临床上重要的革兰氏阳性菌也显示出强效抑制活性（最低抑菌浓度为2 - 4 mg/L）。此外，与对细菌细胞的作用相比，2d对人宫颈癌细胞的抑制作用显著降低。通过共聚焦显微镜对GFP - FtsZ进行活细胞成像，发现2d能迅速扰乱枯草芽孢杆菌细胞中的细胞分裂FtsZ环。FtsZ的免疫荧光成像也表明，2d在5分钟内就能破坏细菌中的Z环。用2d长时间处理会产生丝状细菌，但2d对细菌的类核或膜电位均无明显影响。2d在体外抑制FtsZ组装，而对微管蛋白组装影响极小。有趣的是，2d强烈增强FtsZ的GTP酶活性并降低微管蛋白的GTP酶活性。此外，2d与纯化的FtsZ结合，解离常数为4.0 ± 1.1 μM，2d的结合改变了FtsZ的二级结构。这些结果共同表明，非天然姜黄素类似物2d对重要病原菌具有强大的抗菌活性，且有证据表明2d通过靶向FtsZ来抑制细菌增殖。
