Analysis of Structure-Function Relationships in the Colibactin-Maturating Enzyme ClbP. - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Journal of Molecular Biology Année : 2012

Analysis of Structure-Function Relationships in the Colibactin-Maturating Enzyme ClbP.

A Cougnoux
  • Fonction : Auteur
L Gibold
  • Fonction : Auteur
F Robin
D Dubois
  • Fonction : Auteur
Nathalie Pradel
A Darfeuille-Michaud
  • Fonction : Auteur
G Dalmasso
  • Fonction : Auteur
J Delmas
R Bonnet
  • Fonction : Auteur

Résumé

pks genomic island of Escherichia coli is involved in the synthesis of the non-ribosomal peptide-type genotoxin colibactin, which has been suggesting as affecting the host immune response and having an impact on cancer development. The pks-encoded enzyme ClbP is an atypical peptidase that contributes to the synthesis of colibactin. In this work, we identified key features of ClbP. Bacterial fractionation and Western-blot analysis revealed the docking of ClbP to the bacterial inner membrane via a C-terminal domain harboring three predicted transmembrane helices. Whereas only one helix was necessary for the location in the inner membrane, the complete sequence of the C-terminal domain was necessary for ClbP bioactivity. In addition, the N-terminal sequence of ClbP allowed the SRP/Sec/YidC- and MreB-dependent translocation of the enzymatic domain in the periplasmic compartment, a feature also essential for ClbP bioactivity. Finally, the comparison of ClbP structure with that of the paralogs FmtA-like and AmpC revealed at an extremity of the catalytic groove a negative electrostatic potential surface characteristic of ClbP. Site-directed mutagenesis experiments identified in this zone two aspartic residues that were important for ClbP bioactivity. Overall, these results suggest a model for precolibactin activation by ClbP and pave a way for the design of inhibitors targeting colibactin production.

Dates et versions

hal-00748966 , version 1 (06-11-2012)

Identifiants

Citer

A Cougnoux, L Gibold, F Robin, D Dubois, Nathalie Pradel, et al.. Analysis of Structure-Function Relationships in the Colibactin-Maturating Enzyme ClbP.. Journal of Molecular Biology, 2012, 424 (3-4), in press. ⟨10.1016/j.jmb.2012.09.017⟩. ⟨hal-00748966⟩
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