Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Mumbai, India.
Biochemistry. 2012 Jul 10;51(27):5434-42. doi: 10.1021/bi201813u. Epub 2012 Jun 28.
A perturbation of FtsZ assembly dynamics has been shown to inhibit bacterial cytokinesis. In this study, the antibacterial activity of 151 rhodanine compounds was assayed using Bacillus subtilis cells. Of 151 compounds, eight strongly inhibited bacterial proliferation at 2 μM. Subsequently, we used the elongation of B. subtilis cells as a secondary screen to identify potential FtsZ-targeted antibacterial agents. We found that three compounds significantly increased bacterial cell length. One of the three compounds, namely, CCR-11 [(E)-2-thioxo-5-({[3-(trifluoromethyl)phenyl]furan-2-yl}methylene)thiazolidin-4-one], inhibited the assembly and GTPase activity of FtsZ in vitro. CCR-11 bound to FtsZ with a dissociation constant of 1.5 ± 0.3 μM. A docking analysis indicated that CCR-11 may bind to FtsZ in a cavity adjacent to the T7 loop and that short halogen-oxygen, H-bonding, and hydrophobic interactions might be important for the binding of CCR-11 with FtsZ. CCR-11 inhibited the proliferation of B. subtilis cells with a half-maximal inhibitory concentration (IC(50)) of 1.2 ± 0.2 μM and a minimal inhibitory concentration of 3 μM. It also potently inhibited proliferation of Mycobacterium smegmatis cells. Further, CCR-11 perturbed Z-ring formation in B. subtilis cells; however, it neither visibly affected nucleoid segregation nor altered the membrane integrity of the cells. CCR-11 inhibited HeLa cell proliferation with an IC(50) value of 18.1 ± 0.2 μM (∼15 × IC(50) of B. subtilis cell proliferation). The results suggested that CCR-11 inhibits bacterial cytokinesis by inhibiting FtsZ assembly, and it can be used as a lead molecule to develop FtsZ-targeted antibacterial agents.
FtsZ 组装动力学的扰动已被证明能抑制细菌胞质分裂。在这项研究中，使用枯草芽孢杆菌细胞测定了 151 种罗哚宁化合物的抗菌活性。在 151 种化合物中，有 8 种在 2 μM 时强烈抑制细菌增殖。随后，我们使用枯草芽孢杆菌细胞的伸长作为二次筛选来鉴定潜在的 FtsZ 靶向抗菌剂。我们发现三种化合物显著增加了细菌细胞长度。这三种化合物之一，即 CCR-11 [(E)-2-硫代-5-([[3-(三氟甲基)苯基]呋喃-2-基]亚甲基)噻唑烷-4-酮]，体外抑制 FtsZ 的组装和 GTPase 活性。CCR-11 与 FtsZ 的解离常数为 1.5 ± 0.3 μM。对接分析表明，CCR-11 可能结合在 T7 环附近的 FtsZ 腔中，短卤氧、氢键和疏水性相互作用可能对 CCR-11 与 FtsZ 的结合很重要。CCR-11 抑制枯草芽孢杆菌细胞的增殖，半数最大抑制浓度（IC（50））为 1.2 ± 0.2 μM，最小抑制浓度为 3 μM。它还能有效抑制耻垢分枝杆菌细胞的增殖。此外，CCR-11 扰乱了枯草芽孢杆菌细胞的 Z 环形成；然而，它既没有明显影响核区分离，也没有改变细胞的膜完整性。CCR-11 抑制 HeLa 细胞增殖，IC（50）值为 18.1 ± 0.2 μM（枯草芽孢杆菌细胞增殖的约 15×IC（50））。结果表明，CCR-11 通过抑制 FtsZ 组装抑制细菌胞质分裂，可作为开发 FtsZ 靶向抗菌剂的先导分子。
