title: Plumbagin inhibits cytokinesis in Bacillus subtilis by inhibiting FtsZ assembly--a mechanistic study of its antibacterial activity.
abstract: The assembly of FtsZ plays a central role in construction of the cytokinetic Z-ring that orchestrates bacterial cell division. A naturally occurring naphthoquinone, plumbagin, is known to exhibit antibacterial properties against several types of bacteria. In this study, plumbagin was found to perturb formation of the Z-ring in Bacillus subtilis 168 cells and to cause elongation of these cells without an apparent effect on nucleoid segregation, indicating that it may inhibit FtsZ assembly. Furthermore, it bound to purified B. subtilis FtsZ (BsFtsZ) with a dissociation constant of 20.7 ± 5.6 μM, and inhibited the assembly and GTPase activity of BsFtsZ in vitro. Interestingly, plumbagin did not inhibit either the assembly or GTPase activity of Escherichia coli FtsZ (EcFtsZ) in vitro. Using docking analysis, a putative plumbagin-binding site on BsFtsZ was identified, and the analysis indicated that hydrophobic interactions and hydrogen bonds predominate. Based on the in silico analysis, two variants of BsFtsZ, namely D199A and V307R, were constructed to explore the binding interaction of plumbagin and BsFtsZ. The effects of plumbagin on the assembly and GTPase activity of the variant BsFtsZ proteins in vitro indicated that the residues D199 and V307 may be involved in the binding of plumbagin to BsFtsZ. The results suggest that plumbagin inhibits bacterial proliferation by inhibiting the assembly of FtsZ, and provide insight into the binding site of plumbagin on BsFtsZ, which may help in the design of potent FtsZ-targeted antibacterial agents.
authors.affiliation: Department of Chemistry, Indian Institute of Technology Bombay, Mumbai, India.
authors.forename: Anusri
authors.initials: A
authors.lastname: Bhattacharya
authors.forename: Bhavya
authors.initials: B
authors.lastname: Jindal
authors.forename: Parminder
authors.initials: P
authors.lastname: Singh
authors.forename: Anindya
authors.initials: A
authors.lastname: Datta
authors.forename: Dulal
authors.initials: D
authors.lastname: Panda
mesh_terms: D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology*; D001224:Aspartic Acid / Q000737:chemistry / Q000235:genetics; D001412:Bacillus subtilis / Q000737:chemistry / Q000187:drug effects* / Q000235:genetics; D058491:Bacterial Load; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics; D001665:Binding Sites; D048749:Cytokinesis / Q000187:drug effects* / Q000235:genetics; D003598:Cytoskeletal Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics; D004926:Escherichia coli / Q000737:chemistry / Q000187:drug effects / Q000235:genetics; D006860:Hydrogen Bonding; D057927:Hydrophobic and Hydrophilic Interactions; D062105:Molecular Docking Simulation; D009154:Mutation; D009285:Naphthoquinones / Q000737:chemistry / Q000494:pharmacology*; D011485:Protein Binding; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics; D013045:Species Specificity; D014633:Valine / Q000737:chemistry / Q000235:genetics
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D003598:Cytoskeletal Proteins; C071525:FtsZ protein, Bacteria; D009285:Naphthoquinones; D011994:Recombinant Proteins; D001224:Aspartic Acid; D014633:Valine; C014758:plumbagin
keywords: FtsZ assembly; Z-ring; antibacterial agent; bacterial cell division; plumbagin