Department of Chemistry and Institute of Natural Science, Hanyang University, Seoul, 133-791, Republic of Korea.
Appl Biochem Biotechnol. 2014 Mar;172(6):3263-70. doi: 10.1007/s12010-014-0752-2. Epub 2014 Feb 9.
FtsZ is an essential bacterial cell division protein that is an attractive target for the development of antibacterial agents. FtsZ is a homologue of eukaryotic tubulin, has GTPase activity, and forms a ring-type structure to initiate cell division. In this study, the FtsZ of Bacillus anthracis was cloned into a bacterial expression vector and overexpressed into Escherichia coli BL21 (DE3) cells. The overexpressed B. anthracis FtsZ was soluble and purified to homogeneity using Ni-His-tag affinity chromatography. Like other known FtsZs, the recombinant B. anthracis FtsZ also showed GTP-dependent polymerization, which was analyzed using both spectrophotometric and Transmission Electronic Microscopic (TEM) analysis. Using the purified FtsZ, we screened a naturally extracted chemical library to identify potent and novel inhibitors. The screening yielded three chemicals, SA-011, SA-059, and SA-069, that inhibited the in vitro polymerization activity of FtsZ in the micromolar range (IC50 of 55-168 μM). The inhibition potency was significantly comparable with that of berberine, a known potential inhibitor of FtsZ. Understanding the biochemical basis of the effect of these inhibitors on B. anthracis growth would provide a promising path for the development of new antianthracis drugs.
FtsZ 是一种重要的细菌细胞分裂蛋白，是开发抗菌药物的有吸引力的靶标。FtsZ 是真核微管蛋白的同源物，具有 GTPase 活性，并形成环形结构以启动细胞分裂。在这项研究中，炭疽杆菌的 FtsZ 被克隆到细菌表达载体中，并在大肠杆菌 BL21（DE3）细胞中过表达。使用 Ni-His 标签亲和层析对过表达的 B. anthracis FtsZ 进行可溶性和均相纯化。与其他已知的 FtsZ 一样，重组 B. anthracis FtsZ 还表现出 GTP 依赖性聚合，这通过分光光度法和透射电子显微镜（TEM）分析进行了分析。使用纯化的 FtsZ，我们从天然提取的化学文库中筛选出潜在的新型抑制剂。筛选出三种化学物质，SA-011、SA-059 和 SA-069，它们在微摩尔范围内抑制 FtsZ 的体外聚合活性（IC50 为 55-168 μM）。抑制效力与已知的 FtsZ 潜在抑制剂黄连素相当。了解这些抑制剂对炭疽杆菌生长的生化基础将为开发新型抗炭疽药物提供有希望的途径。
