title: The outer-membrane export signal of Porphyromonas gingivalis type IX secretion system (T9SS) is a conserved C-terminal β-sandwich domain.
abstract: In the recently characterized Type IX Secretion System (T9SS), the conserved C-terminal domain (CTD) in secreted proteins functions as an outer membrane translocation signal for export of virulence factors to the cell surface in the Gram-negative Bacteroidetes phylum. In the periodontal pathogen Porphyromonas gingivalis, the CTD is cleaved off by PorU sortase in a sequence-independent manner, and anionic lipopolysaccharide (A-LPS) is attached to many translocated proteins, thus anchoring them to the bacterial surface. Here, we solved the atomic structure of the CTD of gingipain B (RgpB) from P. gingivalis, alone and together with a preceding immunoglobulin-superfamily domain (IgSF). The CTD was found to possess a typical Ig-like fold encompassing seven antiparallel β-strands organized in two β-sheets, packed into a β-sandwich structure that can spontaneously dimerise through C-terminal strand swapping. Small angle X-ray scattering (SAXS) revealed no fixed orientation of the CTD with respect to the IgSF. By introducing insertion or substitution of residues within the inter-domain linker in the native protein, we were able to show that despite the region being unstructured, it nevertheless is resistant to general proteolysis. These data suggest structural motifs located in the two adjacent Ig-like domains dictate the processing of CTDs by the T9SS secretion pathway.
authors.affiliation: Proteolysis Lab; Department of Structural Biology; "María de Maeztu" Unit of Excellence; Molecular Biology Institute of Barcelona, CSIC; Barcelona Science Park, Barcelona, Spain.
authors.forename: Iñaki
authors.initials: I
authors.lastname: de Diego
authors.affiliation: Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.|Małopolska Centre of Biotechnology, Jagiellonian University, Kraków, Poland.|Department of Oral Immunology and Infectious Diseases, University of Louisville School of Dentistry, Louisville, KY, USA.
authors.forename: Miroslaw
authors.initials: M
authors.lastname: Ksiazek
authors.affiliation: Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.
authors.forename: Danuta
authors.initials: D
authors.lastname: Mizgalska
authors.affiliation: Department of Oral Immunology and Infectious Diseases, University of Louisville School of Dentistry, Louisville, KY, USA.
authors.forename: Lahari
authors.initials: L
authors.lastname: Koneru
authors.affiliation: Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.
authors.forename: Przemyslaw
authors.initials: P
authors.lastname: Golik
authors.affiliation: Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.
authors.forename: Borys
authors.initials: B
authors.lastname: Szmigielski
authors.affiliation: Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Nowak
authors.affiliation: Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.
authors.forename: Zuzanna
authors.initials: Z
authors.lastname: Nowakowska
authors.affiliation: Department of Oral Immunology and Infectious Diseases, University of Louisville School of Dentistry, Louisville, KY, USA.
authors.forename: Barbara
authors.initials: B
authors.lastname: Potempa
authors.affiliation: Department of Oral Immunology and Infectious Diseases, University of Louisville School of Dentistry, Louisville, KY, USA.
authors.forename: John A
authors.initials: JA
authors.lastname: Houston
authors.affiliation: Department of Molecular Biology and Genetics and Interdisciplinary Nanoscience Center, Aarhus University, Aarhus, Denmark.
authors.forename: Jan J
authors.initials: JJ
authors.lastname: Enghild
authors.affiliation: Department of Molecular Biology and Genetics and Interdisciplinary Nanoscience Center, Aarhus University, Aarhus, Denmark.
authors.forename: Ida B
authors.initials: IB
authors.lastname: Thøgersen
authors.affiliation: Faculty of Dentistry, University of Sydney, Sydney, Australia.|Institute of Dental Research, Westmead Centre for Oral Health and Westmead Institute for Medical Research, Sydney, Australia.
authors.forename: Jinlong
authors.initials: J
authors.lastname: Gao
authors.affiliation: Faculty of Science, University of Sydney, Sydney, Australia.
authors.forename: Ann H
authors.initials: AH
authors.lastname: Kwan
authors.affiliation: Faculty of Science, University of Sydney, Sydney, Australia.
authors.forename: Jill
authors.initials: J
authors.lastname: Trewhella
authors.affiliation: Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.|Małopolska Centre of Biotechnology, Jagiellonian University, Kraków, Poland.
authors.forename: Grzegorz
authors.initials: G
authors.lastname: Dubin
authors.affiliation: Proteolysis Lab; Department of Structural Biology; "María de Maeztu" Unit of Excellence; Molecular Biology Institute of Barcelona, CSIC; Barcelona Science Park, Barcelona, Spain.
authors.forename: F Xavier
authors.initials: FX
authors.lastname: Gomis-Rüth
authors.affiliation: Faculty of Dentistry, University of Sydney, Sydney, Australia.|Institute of Dental Research, Westmead Centre for Oral Health and Westmead Institute for Medical Research, Sydney, Australia.
authors.forename: Ky-Anh
authors.initials: KA
authors.lastname: Nguyen
authors.affiliation: Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.|Małopolska Centre of Biotechnology, Jagiellonian University, Kraków, Poland.|Department of Oral Immunology and Infectious Diseases, University of Louisville School of Dentistry, Louisville, KY, USA.
authors.forename: Jan
authors.initials: J
authors.lastname: Potempa
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000378:metabolism; D058947:Bacterial Secretion Systems / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D017124:Conserved Sequence; D007136:Immunoglobulins / Q000378:metabolism*; D008958:Models, Molecular; D049790:Nuclear Export Signals / Q000235:genetics*; D016966:Porphyromonas gingivalis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017433:Protein Structure, Secondary; D021381:Protein Transport; D053838:Scattering, Small Angle
chemical_list: D001426:Bacterial Proteins; D058947:Bacterial Secretion Systems; D007136:Immunoglobulins; D049790:Nuclear Export Signals