Sanguinarine blocks cytokinesis in bacteria by inhibiting FtsZ assembly and bundling
Sanguinarine blocks cytokinesis in bacteria by inhibiting FtsZ assembly and bundling
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DOI：
10.1021/bi050767
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发表时间：
2005-12-20
期刊：
影响因子：
2.9
通讯作者：
Panda, D
中科院分区：
文献类型：
--
作者：
Beuria, TK;Santra, MK;Panda, D
细菌性疾病是人类死亡的主要原因之一。抗生素耐药性的发展大大有助于高死亡率，因此，需要发现具有新作用机制的抗菌药物。在这项研究中，我们发现，血根碱，一个benzophenanthridine生物碱，强烈地诱导在革兰氏阳性和革兰氏阴性细菌和抑制胞质分裂，防止细菌细胞分裂。血根碱没有扰乱大肠杆菌的膜结构。然而，它干扰了E.杆菌此外，血根碱强烈降低了Z环/微米枯草芽孢杆菌长度的发生频率，但没有改变细胞长度的类核数量/微米。结果表明血根碱抑制B细胞的胞质分裂。枯草芽孢杆菌通过抑制Z-环形成而不影响类核分离。血根碱在体外抑制纯化的FtsZ的组装，并减少FtsZ原丝的成束。此外，血根碱与FtsZ的相互作用进行了研究，使用尺寸排阻色谱法，外源性荧光探针1-苯胺基萘-8-磺酸，和色氨酸荧光突变FtsZ（Y371 W）。发现血根碱以18-30 μ M的解离常数与FtsZ结合。结果表明血根碱通过干扰Z环中FtsZ组装动力学来抑制细菌分裂，并提供证据支持FtsZ的组装和捆绑在细菌胞质分裂中起关键作用的假设。结果表明，血根碱可作为先导化合物开发FtsZ靶向抗菌剂。
Bacterial diseases are among the leading causes of human death. The development of antibiotic resistance greatly contributes to the high mortality rate, and thus, the discovery of antibacterial drugs with novel mechanisms of action is needed. In this study, we found that sanguinarine, a benzophenanthridine alkaloid, strongly induced filamentation in both Gram-positive and Gram-negative bacteria and prevented bacterial cell division by inhibiting cytokinesis. Sanguinarine did not perturb the membrane structure in Escherichia coli. However, it perturbed the cytokinetic Z-ring formation in E. coli. In addition, sanguinarine strongly reduced the frequency of the occurrence of Z rings/micrometer of Bacillus subtilis length but did not alter the number of nucleoids/micrometer of cell length. The results suggested that sanguinarine inhibited cytokinesis in B. subtilis by inhibiting Z-ring formation without affecting nucleoid segregation. Sanguinarine inhibited the assembly of purified FtsZ and reduced the bundling of FtsZ protofilaments in vitro. Further, the interaction of sanguinarine to FtsZ was investigated using size-exclusion chromatography, an extrinsic fluorescent probe 1-anilinonaphthalene-8-sulfonic acid, and tryptophan fluorescence of mutated FtsZ (Y371W). Sanguinarine was found to bind to FtsZ with a dissociation constant of 18-30 mu M. The results together show that sanguinarine inhibits bacterial division by perturbing FtsZ assembly dynamics in the Z ring and provide evidence in Support of the hypothesis that the assembly and bundling of FtsZ play a critical role in bacterial cytokinesis. The results suggest that sanguinarine may be used as a lead compound to develop FtsZ-targeted antibacterial agents.