title: Structural Basis of Lipid Targeting and Destruction by the Type V Secretion System of Pseudomonas aeruginosa.
abstract: The type V secretion system is a macromolecular machine employed by a number of bacteria to secrete virulence factors into the environment. The human pathogen Pseudomonas aeruginosa employs the newly described type Vd secretion system to secrete a soluble variant of PlpD, a lipase of the patatin-like family synthesized as a single macromolecule that also carries a polypeptide transport-associated domain and a 16-stranded β-barrel. Here we report the crystal structure of the secreted form of PlpD in its biologically active state. PlpD displays a classical lipase α/β hydrolase fold with a catalytic site located within a highly hydrophobic channel that entraps a lipidic molecule. The active site is covered by a flexible lid, as in other lipases, indicating that this region in PlpD must modify its conformation in order for catalysis at the water-lipid interface to occur. PlpD displays phospholipase A1 activity and is able to recognize a number of phosphatidylinositols and other phosphatidyl analogs. PlpD is the first example of an active phospholipase secreted through the type V secretion system, for which there are more than 200 homologs, revealing details of the lipid destruction arsenal expressed by P. aeruginosa in order to establish infection.
authors.affiliation: Brazilian National Laboratory for Biosciences (LNBio), CNPEM, Campinas, São Paulo, Brazil.
authors.forename: Paulo Vinicius
authors.initials: PV
authors.lastname: da Mata Madeira
authors.affiliation: Brazilian National Laboratory for Biosciences (LNBio), CNPEM, Campinas, São Paulo, Brazil.
authors.forename: Samira
authors.initials: S
authors.lastname: Zouhir
authors.affiliation: Univ. Grenoble Alpes, Bacterial Pathogenesis and Cellular Responses Group, Institut de Biosciences et Biotechnologies de Grenoble (BIG), Grenoble, France; CNRS, IBS and BIG, Grenoble, France; CEA, IBS and BIG, Grenoble, France; INSERM, UMR-S 1036 Biology of Cancer and Infection Laboratory, Grenoble, France.
authors.forename: Pauline
authors.initials: P
authors.lastname: Basso
authors.affiliation: Brazilian National Laboratory for Biosciences (LNBio), CNPEM, Campinas, São Paulo, Brazil.
authors.forename: David
authors.initials: D
authors.lastname: Neves
authors.affiliation: CNRS & Aix-Marseille Université, Laboratoire d'Ingénierie des Systèmes Macromoléculaires (UMR7255), Institut de Microbiologie de la Méditerranée, Marseille, France.
authors.forename: Aurélie
authors.initials: A
authors.lastname: Laubier
authors.affiliation: CNRS & Aix-Marseille Université, Laboratoire d'Ingénierie des Systèmes Macromoléculaires (UMR7255), Institut de Microbiologie de la Méditerranée, Marseille, France.
authors.forename: Richard
authors.initials: R
authors.lastname: Salacha
authors.affiliation: CNRS & Aix-Marseille Université, Laboratoire d'Ingénierie des Systèmes Macromoléculaires (UMR7255), Institut de Microbiologie de la Méditerranée, Marseille, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Bleves
authors.affiliation: Univ. Grenoble Alpes, Bacterial Pathogenesis and Cellular Responses Group, Institut de Biosciences et Biotechnologies de Grenoble (BIG), Grenoble, France; CNRS, IBS and BIG, Grenoble, France; CEA, IBS and BIG, Grenoble, France; INSERM, UMR-S 1036 Biology of Cancer and Infection Laboratory, Grenoble, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Faudry
authors.affiliation: CNRS, IBS and BIG, Grenoble, France; CEA, IBS and BIG, Grenoble, France; Univ. Grenoble Alpes, Institut de Biologie Structurale (IBS), Grenoble, France. Electronic address: carlos.contreras@ibs.fr.
authors.forename: Carlos
authors.initials: C
authors.lastname: Contreras-Martel
authors.affiliation: Brazilian National Laboratory for Biosciences (LNBio), CNPEM, Campinas, São Paulo, Brazil; CNRS, IBS and BIG, Grenoble, France; CEA, IBS and BIG, Grenoble, France; Univ. Grenoble Alpes, Institut de Biologie Structurale (IBS), Grenoble, France. Electronic address: andrea.dessen@ibs.fr.
authors.forename: Andréa
authors.initials: A
authors.lastname: Dessen
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism*; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D008958:Models, Molecular; D010716:Phosphatidylinositols / Q000378:metabolism; D010740:Phospholipases / Q000737:chemistry* / Q000378:metabolism*; D011487:Protein Conformation; D011550:Pseudomonas aeruginosa / Q000201:enzymology*; D013379:Substrate Specificity; D000069320:Type V Secretion Systems / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D010716:Phosphatidylinositols; D000069320:Type V Secretion Systems; D010740:Phospholipases
keywords: bacterial secretion; crystallography; infection; lipid affinity; phospholipase