Systematic mutational analysis of the putative hydrolase PqsE: toward a deeper molecular understanding of virulence acquisition in Pseudomonas aeruginosa
| Autor: | Eric Déziel, Benjamin Folch, Nicolas Doucet |
|---|---|
| Přispěvatelé: | Institut Armand Frappier (INRS-IAF), Réseau International des Instituts Pasteur (RIIP)-Institut National de la Recherche Scientifique [Québec] (INRS), Groupe de Recherche Axé sur la Structure des Protéines (GRASP), McGill University = Université McGill [Montréal, Canada], PROTEO, The Quebec Network for Research on Protein Function, Engineering, and Applications, Réseau International des Instituts Pasteur (RIIP)-Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Recherche Scientifique [Québec] (INRS)-Université de Sherbrooke (UdeS)-Université Laval [Québec] (ULaval)-McGill University = Université McGill [Montréal, Canada]-University of Ottawa [Ottawa]-Université du Québec à Trois-Rivières (UQTR)-Université de Montréal (UdeM)-TransBiotech, Lévis-Concordia University [Montreal]-Université du Québec à Montréal = University of Québec in Montréal (UQAM), This work was supported by Natural Sciences and Engineering Research Council Discovery grant RGPIN 402623-2011 (to N.D.), CIHR operating grant MOP-97888 (to E.D.), and a FRQS Research Scholar Junior 1 Career Award (to N.D.). N.D. also acknowledges support from the FRQNT Strategic Cluster 'Regroupement Québécois de Recherche sur la Fonction, la Structure et l'Ingénierie des Protéines' (PROTEO) and the FRQS Strategic Cluster 'Groupe de Recherche Axé sur la Structure des Protéines' (GRASP). E.D. holds a Canada Research Chair in sociomicrobiology. B.F. is the recipient of a 'Fondation Universitaire Armand-Frappier de l'INRS' postdoctoral fellowship. |
| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
MESH: Pseudomonas aeruginosa/metabolism
Models Molecular MESH: Hydrolases/genetics Protein Folding MESH: Pseudomonas Infections/microbiology Operon MESH: Sequence Homology Amino Acid Hydrolases [SDV]Life Sciences [q-bio] MESH: Protein Structure Secondary lcsh:Medicine MESH: Amino Acid Sequence medicine.disease_cause Protein Structure Secondary chemistry.chemical_compound MESH: Protein Structure Tertiary lcsh:Science Peptide sequence Genetics 0303 health sciences Multidisciplinary Virulence 030302 biochemistry & molecular biology MESH: Pseudomonas aeruginosa/genetics Alanine scanning MESH: Hydrolases/metabolism Pseudomonas aeruginosa Thermodynamics MESH: Thermodynamics MESH: Bacterial Proteins/metabolism MESH: Models Molecular MESH: Bacterial Proteins/chemistry Research Article MESH: Mutation MESH: Pseudomonas aeruginosa/pathogenicity MESH: Protein Folding Molecular Sequence Data Biology MESH: Bacterial Proteins/genetics MESH: Mutagenesis Site-Directed/methods 03 medical and health sciences Pyocyanin Bacterial Proteins Hydrolase MESH: Hydrolases/chemistry medicine Humans [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Pseudomonas Infections Amino Acid Sequence MESH: Virulence/genetics 030304 developmental biology MESH: Pyocyanine/metabolism MESH: Molecular Sequence Data MESH: Humans Binding Sites MESH: Binding Sites/genetics Sequence Homology Amino Acid lcsh:R Protein Structure Tertiary Quorum sensing chemistry Mutation Mutagenesis Site-Directed Pyocyanine lcsh:Q |
| Zdroj: | PLoS ONE, Vol 8, Iss 9, p e73727 (2013) PLoS ONE PLoS ONE, Public Library of Science, 2013, 8 (9), pp.e73727. ⟨10.1371/journal.pone.0073727⟩ |
| ISSN: | 1932-6203 |
| Popis: | International audience; Pseudomonas aeruginosa is an important opportunistic human pathogen that can establish bacterial communication by synchronizing the behavior of individual cells in a molecular phenomenon known as "quorum sensing". Through an elusive mechanism involving gene products of the pqs operon, the PqsE enzyme is absolutely required for the synthesis of extracellular phenazines, including the toxic blue pigment pyocyanin, effectively allowing cells to achieve full-fledged virulence. Despite several functional and structural attempts at deciphering the role of this relevant enzymatic drug target, no molecular function has yet been ascribed to PqsE. In the present study, we report a series of alanine scanning experiments aimed at altering the biological function of PqsE, allowing us to uncover key amino acid positions involved in the molecular function of this enzyme. We use sequence analysis and structural overlays with members of homologous folds to pinpoint critical positions located in the vicinity of the ligand binding cleft and surrounding environment, revealing the importance of a unique C-terminal α-helical motif in the molecular function of PqsE. Our results suggest that the active site of the enzyme involves residues that extend further into the hydrophobic core of the protein, advocating for a lid-like movement of the two terminal helices. This information should help design virtual libraries of PqsE inhibitors, providing means to counter P. aeruginosa virulence acquisition and helping to reduce nosocomial infections. |
| Databáze: | OpenAIRE |
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