title: Crystal structure of VapC21 from Mycobacterium tuberculosis at 1.31 Å resolution.
abstract: Persisters are individual bacterial cells that exhibit a phenotype characterized by slow growth, low metabolic rate and multidrug tolerance. The processes that drive cells into a persistence state constitute an active but incipient research field, and structural data regarding its components are scarce. The molecular targets of many therapeutic drugs are involved in cell wall synthesis and cell division, and these cellular processes are down-regulated in persister cells, consequently these cells are more likely to survive antibiotic treatment. Toxin-antitoxin systems were shown to have a leading role in the formation of persisters, and several pathogenic bacteria display a wide array of these systems. The Mycobacterium tuberculosis H37Rv genome presents 88 toxin-antitoxin loci, of which 47 code for members of the VapBC protein family. To date, only four crystal structures of Mycobacterium tuberculosis VapBC complexes are available, and all of them present the toxin bound to and inhibited by the antitoxin. We present the 1.31 Å resolution structure of VapC21, the first structure of a Mycobacterium tuberculosis VapC toxin in the absence of its cognate inhibitory antitoxin. Our data show that VapC21 is a dimer in solution, with conserved active site architecture and an extensive antitoxin binding groove. Additionally, the strategy used to mutate a putative catalytic residue allowing the expression and purification of soluble VapC21 will pave the way for the resolution of more toxin structures in the absence of antitoxins. Taken together, our findings represent an important step in unraveling the molecular mechanisms related to persistence, which will contribute for the design of faster and more efficient therapeutic approaches for the treatment of tuberculosis, particularly for infections with multidrug-resistant strains.
authors: Laboratory of Molecular Biophysics, Department of Cell Biology, University of Brasília, Brasília, Brazil.
authors: Paulo
authors: P
authors: Jardim
authors: Laboratory of Molecular Biophysics, Department of Cell Biology, University of Brasília, Brasília, Brazil.
authors: Isadora Cristina da Silva
authors: IC
authors: Santos
authors: Laboratory of Molecular Biophysics, Department of Cell Biology, University of Brasília, Brasília, Brazil.
authors: João Alexandre Ribeiro Gonçalves
authors: JA
authors: Barbosa
authors: Laboratory of Molecular Biophysics, Department of Cell Biology, University of Brasília, Brasília, Brazil.
authors: Sônia Maria
authors: SM
authors: de Freitas
authors: Laboratory of Molecular Biophysics, Department of Cell Biology, University of Brasília, Brasília, Brazil. Electronic address: napo@unb.br.
authors: Napoleão Fonseca
authors: NF
authors: Valadares
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry*; D020134:Catalytic Domain; D017124:Conserved Sequence; D018360:Crystallography, X-Ray; D006860:Hydrogen Bonding; D008958:Models, Molecular; D009169:Mycobacterium tuberculosis / Q000378:metabolism*; D055672:Static Electricity
chemical_list: D001426:Bacterial Proteins
keywords: Crystallography; Heterologous expression; Toxin; VapBC; VapC21