title: Translocation path of a substrate protein through its Omp85 transporter.
abstract: TpsB proteins are Omp85 superfamily members that mediate protein translocation across the outer membrane of Gram-negative bacteria. Omp85 transporters are composed of N-terminal POTRA domains and a C-terminal transmembrane β-barrel. In this work, we track the in vivo secretion path of the Bordetella pertussis filamentous haemagglutinin (FHA), the substrate of the model TpsB transporter FhaC, using site-specific crosslinking. The conserved secretion domain of FHA interacts with the POTRA domains, specific extracellular loops and strands of FhaC and the inner β-barrel surface. The interaction map indicates a funnel-like pathway, with conformationally flexible FHA entering the channel in a non-exclusive manner and exiting along a four-stranded β-sheet at the surface of the FhaC barrel. This sheet of FhaC guides the secretion domain of FHA along discrete steps of translocation and folding. This work demonstrates that the Omp85 barrel serves as a channel for translocation of substrate proteins.
authors.affiliation: 1] Center for Infection and Immunity of Lille, Institut Pasteur de Lille, 1 rue Calmette, Lille 59021, France [2] CNRS UMR8204, Lille 59021, France [3] INSERM U1019, Lille 59045, France [4] University of Lille Nord de France, Lille 59044, France.
authors.forename: Catherine
authors.initials: C
authors.lastname: Baud
authors.affiliation: 1] Center for Infection and Immunity of Lille, Institut Pasteur de Lille, 1 rue Calmette, Lille 59021, France [2] CNRS UMR8204, Lille 59021, France [3] INSERM U1019, Lille 59045, France [4] University of Lille Nord de France, Lille 59044, France.
authors.forename: Jérémy
authors.initials: J
authors.lastname: Guérin
authors.affiliation: 1] Center for Infection and Immunity of Lille, Institut Pasteur de Lille, 1 rue Calmette, Lille 59021, France [2] CNRS UMR8204, Lille 59021, France [3] INSERM U1019, Lille 59045, France [4] University of Lille Nord de France, Lille 59044, France.
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Petit
authors.affiliation: 1] Center for Infection and Immunity of Lille, Institut Pasteur de Lille, 1 rue Calmette, Lille 59021, France [2] CNRS UMR8204, Lille 59021, France [3] INSERM U1019, Lille 59045, France [4] University of Lille Nord de France, Lille 59044, France.
authors.forename: Elodie
authors.initials: E
authors.lastname: Lesne
authors.affiliation: 1] Center for Infection and Immunity of Lille, Institut Pasteur de Lille, 1 rue Calmette, Lille 59021, France [2] CNRS UMR8204, Lille 59021, France [3] INSERM U1019, Lille 59045, France [4] University of Lille Nord de France, Lille 59044, France.
authors.forename: Elian
authors.initials: E
authors.lastname: Dupré
authors.affiliation: 1] Center for Infection and Immunity of Lille, Institut Pasteur de Lille, 1 rue Calmette, Lille 59021, France [2] CNRS UMR8204, Lille 59021, France [3] INSERM U1019, Lille 59045, France [4] University of Lille Nord de France, Lille 59044, France.
authors.forename: Camille
authors.initials: C
authors.lastname: Locht
authors.affiliation: 1] Center for Infection and Immunity of Lille, Institut Pasteur de Lille, 1 rue Calmette, Lille 59021, France [2] CNRS UMR8204, Lille 59021, France [3] INSERM U1019, Lille 59045, France [4] University of Lille Nord de France, Lille 59044, France.
authors.forename: Françoise
authors.initials: F
authors.lastname: Jacob-Dubuisson
mesh_terms: D018829:Adhesins, Bacterial / Q000378:metabolism*; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D001886:Bordetella pertussis / Q000378:metabolism*; D003432:Cross-Linking Reagents / Q000737:chemistry; D003545:Cysteine / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism; D005434:Flow Cytometry; D015964:Gene Expression Regulation, Bacterial; D006388:Hemagglutinins / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D013379:Substrate Specificity; D010566:Virulence Factors, Bordetella / Q000378:metabolism*
chemical_list: D018829:Adhesins, Bacterial; D001425:Bacterial Outer Membrane Proteins; D003432:Cross-Linking Reagents; D006388:Hemagglutinins; D026901:Membrane Transport Proteins; D010446:Peptide Fragments; D010566:Virulence Factors, Bordetella; C098052:fhaC protein, Bordetella pertussis; C065273:filamentous hemagglutinin adhesin, Bordetella pertussis; D003545:Cysteine