Totarol inhibits bacterial cytokinesis by perturbing the assembly dynamics of FtsZ
Totarol inhibits bacterial cytokinesis by perturbing the assembly dynamics of FtsZ
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DOI：
10.1021/bi602573e
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发表时间：
2007-04-10
期刊：
影响因子：
2.9
通讯作者：
Panda, Dulal
中科院分区：
文献类型：
--
作者：
Jaiswal, Richa;Beuria, Tushar K.;Panda, Dulal
Totarol是一种二萜类酚，已被证明可抑制几种致病性革兰氏阳性菌（包括结核分枝杆菌）的增殖。在这项研究中，发现totarol抑制枯草芽孢杆菌细胞的增殖，最小抑制浓度为2 μ M。未检测到B对B的膜结构的干扰。枯草芽孢杆菌（B. subtilis）中，它强烈诱导了B的表达。枯草杆菌细胞，这表明它抑制细菌胞质分裂。Totarol（1.5 μ M）降低了每微米细菌细胞长度的Z环出现频率，但不影响类核频率，这表明它通过抑制Z环的形成来阻断胞质分裂。FtsZ的组装动力学被认为在Z环的形成和功能中发挥重要作用，Z环是一种设计细菌胞质分裂的机器。由于妥他洛尔能抑制M.结核病，我们研究了Totarol对M.结核FtsZ（MtbFtsZ）的体外表达。Totarol减少了MtbFtsZ原丝的组装，并有效地抑制了MtbFtsZ的GT酶活性。它与MtbFtsZ结合，解离常数为11 +/- 2.3 μ M。它增加了MtbFtsZ-1-苯胺基萘-8-磺酸复合物的荧光强度，并抑制了N-（1-芘）马来酰亚胺标记的MtbFtsZ的荧光强度，表明妥他洛尔诱导MtbFtsZ的构象变化。结果表明，totarol可以扰乱FtsZ原丝在Z环中的组装动力学。Totarol对HeLa细胞增殖的抑制作用极弱。它不影响HeLa细胞中的微管组织。结果表明，Totarol通过靶向FtsZ抑制细菌增殖，它可能是有用的先导化合物，以开发一种有效的抗结核药物。
Totarol, a diterpenoid phenol, has been shown to inhibit the proliferation of several pathogenic Gram-positive bacteria including Mycobacterium tuberculosis. In this study, totarol was found to inhibit the proliferation of Bacillus subtilis cells with a minimum inhibitory concentration of 2 mu M. It did not detectably perturb the membrane structure of B. subtilis; it strongly induced the filamentation in B. subtilis cells, suggesting that it inhibits bacterial cytokinesis. Totarol (1.5 mu M) reduced the frequency of the Z-ring occurrence per micrometer of the bacterial cell length but did not affect the nucleoid frequency, suggesting that it blocks cytokinesis by inhibiting the formation of the Z-ring. The assembly dynamics of FtsZ is thought to play an important role in the formation and functioning of the Z-ring, a machine that engineers cytokinesis in bacteria. Since totarol was shown to inhibit the proliferation of M. tuberculosis, we examined the effects of totarol on the assembly dynamics of M. tuberculosis FtsZ (MtbFtsZ) in vitro. Totarol decreased the assembly of MtbFtsZ protofilaments and potently suppressed the GTPase activity of MtbFtsZ. It bound to MtbFtsZ with a dissociation constant of 11 +/- 2.3 mu M. It increased the fluorescence intensity of the MtbFtsZ-1-anilinonaphthalene-8-sulfonic acid complex and inhibited the fluorescence intensity of N-(1-pyrene)maleimide-labeled MtbFtsZ, suggesting that totarol induces conformational changes in MtbFtsZ. The results indicated that totarol can perturb the assembly dynamics of FtsZ protofilaments in the Z-ring. Totarol exhibited extremely weak inhibitory effects on HeLa cell proliferation. It did not affect microtubule organization in HeLa cells. The results suggest that totarol inhibits bacterial proliferation by targeting FtsZ and it may be useful as a lead compound to develop an effective antitubercular drug.