Person: Andreu Rodríguez, José Manuel
Universidad Complutense de Madrid
Faculty / Institute
Personalidad, Evaluación y Psicología Clínica
Personalidad, Evaluación y Tratamiento Psicológico
Now showing 1 - 2 of 2
PublicationThe structural assembly switch of cell division protein FtsZ probed with fluorescent allosteric inhibitors(RSC, 2017) Artola Pérez de Azanza, Marta Elena; Ruiz Ávila, Laura, Laura B.; Ramírez Aportela, Erney; Fernando Martínez, R.; Araujo Bazán, Lidia; Vázquez Villa, Henar; Martín Fontecha, María del Mar; Oliva Blanco, María Ángela; Martín Galiano, Antonio Javier; Chacón Montes, Pablo; López Rodríguez, María Luz; Andreu Rodríguez, José Manuel; Huecas Gayo, SoniaFtsZ is a widely conserved tubulin-like GTPase that directs bacterial cell division and a new target for antibiotic discovery. This protein assembly machine cooperatively polymerizes forming single-stranded filaments, by means of self-switching between inactive and actively associating monomer conformations. The structural switch mechanism was proposed to involve a movement of the C-terminal and N-terminal FtsZ domains, opening a cleft between them, allosterically coupled to the formation of a tight association interface between consecutive subunits along the filament. The effective antibacterial benzamide PC190723 binds into the open interdomain cleft and stabilizes FtsZ filaments, thus impairing correct formation of the FtsZ ring for cell division. We have designed fluorescent analogs of PC190723 to probe the FtsZ structural assembly switch. Among them, nitrobenzoxadiazole probes specifically bind to assembled FtsZ rather than to monomers. Probes with several spacer lengths between the fluorophore and benzamide moieties suggest a binding site extension along the interdomain cleft. These probes label FtsZ rings of live Bacillus subtilis and Staphylococcus aureus, without apparently modifying normal cell morphology and growth, but at high concentrations they induce impaired bacterial division phenotypes typical of benzamide antibacterials. During the FtsZ assembly-disassembly process, the fluorescence anisotropy of the probes changes upon binding and dissociating from FtsZ, thus reporting open and closed FtsZ interdomain clefts. Our results demonstrate the structural mechanism of the FtsZ assembly switch, and suggest that the probes bind into the open clefts in cellular FtsZ polymers preferably to unassembled FtsZ in the bacterial cytosol. PublicationTargeting the FtsZ Allosteric Binding Site with a Novel Fluorescence Polarization Screen, Cytological and Structural Approaches for Antibacterial Discovery(ACS Publications, 2021-04-28) Huecas , Sonia; Araújo Bazán , Lidia; Rúiz, Federico M.; Ruiz Ávila, , Laura B.; Martínez, R.Fernando; Escobar Peña, Ana Andrea; Artola, Marta; Vázquez Villa, María Del Henar; Martín-Fontecha Corrales, María Del Mar; Fernández Tornero, Carlos; López Rodríguez, María L.; Andreu Rodríguez, José ManuelBacterial resistance to antibiotics makes previously manageable infections again disabling and lethal, highlighting the need for new antibacterial strategies. In this regard, inhibition of the bacterial division process by targeting key protein FtsZ has been recognized as an attractive approach for discovering new antibiotics. Binding of small molecules to the cleft between the N-terminal guanosine triphosphate (GTP)-binding and the C-terminal subdomains allosterically impairs the FtsZ function, eventually inhibiting bacterial division. Nonetheless, the lack of appropriate chemical tools to develop a binding screen against this site has hampered the discovery of FtsZ antibacterial inhibitors. Herein, we describe the first competitive binding assay to identify FtsZ allosteric ligands interacting with the interdomain cleft, based on the use of specific high-affinity fluorescent probes. This novel assay, together with phenotypic profiling and X-ray crystallographic insights, enables the identification and characterization of FtsZ inhibitors of bacterial division aiming at the discovery of more effective antibacterials.