title: Characterization and in vitro inhibition studies of Bacillus anthracis FtsZ: a potential antibacterial target.
abstract: 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.
authors: Department of Chemistry and Institute of Natural Science, Hanyang University, Seoul, 133-791, Republic of Korea.
authors: Hae-Chul
authors: HC
authors: Park
authors: Vinayakumar
authors: V
authors: Gedi
authors: June-Haeng
authors: JH
authors: Cho
authors: Jae-Wook
authors: JW
authors: Hyun
authors: Kwang-Jick
authors: KJ
authors: Lee
authors: JeongWoo
authors: J
authors: Kang
authors: ByungJae
authors: B
authors: So
authors: Moon-Young
authors: MY
authors: Yoon
mesh_terms: D000900:Anti-Bacterial Agents / Q000737:chemistry*; D001408:Bacillus anthracis / Q000737:chemistry* / Q000201:enzymology; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D001599:Berberine / Q000737:chemistry; D003598:Cytoskeletal Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006160:Guanosine Triphosphate / Q000737:chemistry; D057166:High-Throughput Screening Assays; D055503:Protein Multimerization; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D054852:Small Molecule Libraries / Q000737:chemistry*
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D003598:Cytoskeletal Proteins; D004791:Enzyme Inhibitors; C071525:FtsZ protein, Bacteria; D011993:Recombinant Fusion Proteins; D054852:Small Molecule Libraries; D001599:Berberine; D006160:Guanosine Triphosphate