CSDB: Search results

Taxonomic group: bacteria / Bacteroidetes (Phylum: Bacteroidetes)
Associated disease: periodontitis [ICD11: DA0C

]

NCBI PubMed ID: 33622730
Publication DOI: 10.1128/mBio.02262-20
Journal NLM ID: 101519231
Publisher: Washington, DC: American Society for Microbiology
Correspondence: F. Xavier Gomis-Ruth <xgrcri

ibmb.csic.es>; Jan Potempa <jan.potempa

icloud.com>
Institutions: Department of Microbiology, Faculty of Biochemistry, Biophysics, and Biotechnology, Jagiellonian University, Krakow, Poland, Department of Oral Immunology and Infectious Diseases, University of Louisville School of Dentistry, Louisville, Kentucky, USA, Department of Bacterial Genetics, Institute of Microbiology, Faculty of Biology, University of Warsaw, Warsaw, Poland, Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics, and Biotechnology, Jagiellonian University, Krakow, Poland, Interdisciplinary Nanoscience Center (iNANO) and Department of Molecular Biology, Aarhus University, Aarhus, Denmark, Centre for Host-Microbiome Interactions, Faculty of Dentistry, Oral and Craniofacial Sciences, King's College London, London, United Kingdom, Proteolysis Lab, Department of Structural Biology, Molecular Biology Institute of Barcelona, Higher Scientific Research Council (CSIC), Barcelona, Catalonia, Spain

Cargo proteins of the type IX secretion system (T9SS) in human pathogens from the Bacteroidetes phylum invariably possess a conserved C-terminal domain (CTD) that functions as a signal for outer membrane (OM) translocation. In Porphyromonas gingivalis, the CTD of cargos is cleaved off after translocation, and anionic lipopolysaccharide (A-LPS) is attached. This transpeptidase reaction anchors secreted proteins to the OM. PorZ, a cell surface-associated protein, is an essential component of the T9SS whose function was previously unknown. We recently solved the crystal structure of PorZ and found that it consists of two β-propeller moieties, followed by a CTD. In this study, we performed structure-based modeling, suggesting that PorZ is a carbohydrate-binding protein. Indeed, we found that recombinant PorZ specifically binds A-LPS in vitro Binding was blocked by monoclonal antibodies that specifically react with a phosphorylated branched mannan in the anionic polysaccharide (A-PS) component of A-LPS, but not with the core oligosaccharide or the lipid A endotoxin. Examination of A-LPS derived from a cohort of mutants producing various truncations of A-PS confirmed that the phosphorylated branched mannan is indeed the PorZ ligand. Moreover, purified recombinant PorZ interacted with the PorU sortase in an A-LPS-dependent manner. This interaction on the cell surface is crucial for the function of the "attachment complex" composed of PorU, PorZ, and the integral OM β-barrel proteins PorV and PorQ, which is involved in posttranslational modification and retention of T9SS cargos on the bacterial surface.IMPORTANCE Bacteria have evolved multiple systems to transport effector proteins to their surface or into the surrounding milieu. These proteins have a wide range of functions, including attachment, motility, nutrient acquisition, and toxicity in the host. Porphyromonas gingivalis, the human pathogen responsible for severe gum diseases (periodontitis), uses a recently characterized type IX secretion system (T9SS) to translocate and anchor secreted virulence effectors to the cell surface. Anchorage is facilitated by sortase, an enzyme that covalently attaches T9SS cargo proteins to a unique anionic lipopolysaccharide (A-LPS) moiety of P. gingivalis Here, we show that the T9SS component PorZ interacts with sortase and specifically binds A-LPS. Binding is mediated by a phosphorylated branched mannan repeat in A-LPS polysaccharide. A-LPS-bound PorZ interacts with sortase with significantly higher affinity, facilitating modification of cargo proteins by the cell surface attachment complex of the T9SS.

Lipopolysaccharide, secretion, Porphyromonas gingivalis, gingipains, T9SS

Structure type: oligomer
Location inside paper: Fig. 1
Trivial name: anionic lipopolysaccharide (A-LPS)
Compound class: LPS
Contained glycoepitopes: IEDB_115576,IEDB_120354,IEDB_123890,IEDB_130650,IEDB_130695,IEDB_130701,IEDB_136104,IEDB_137778,IEDB_140116,IEDB_141111,IEDB_141181,IEDB_141793,IEDB_141807,IEDB_141828,IEDB_141829,IEDB_141830,IEDB_141831,IEDB_141832,IEDB_143632,IEDB_144983,IEDB_144996,IEDB_151531,IEDB_152206,IEDB_153220,IEDB_153762,IEDB_153763,IEDB_164174,IEDB_164176,IEDB_474450,IEDB_76933,IEDB_857732,IEDB_857735,IEDB_983930,SB_136,SB_191,SB_196,SB_197,SB_198,SB_44,SB_67,SB_72

Methods: PCR, SDS-PAGE, anion-exchange chromatography, Western blotting, genetic methods, enzymatic digestion, affinity chromatography, SEC, flow cytometry analysis, fluorescence binding assay

NCBI Taxonomy refs (TaxIDs): 837
Show glycosyltransferases

There is only one chemically distinct structure:

CCCCCCCCCCCCCCCC(=O)O[C@H](CCCCCCCCCCCC(C)C)CC(=O)N[C@H]1[C@H](OC[C@H]2O[C@H](OP(=O)(O)O)[C@H](NC(=O)C[C@H](O)CCCCCCCCCCCC(C)C)[C@@H](OC(=O)CC(O)CCCCCCCCCCCCC)[C@@H]2O)O[C@H](CO[C@]2(C(=O)O)C[C@@H](O)[C@@H](OP(=O)(O)OCC(CO[C@H]3O[C@H](CO)[C@@H](O)[C@@H](O)[C@H]3N)O[C@H]3O[C@H](CO)[C@@H](O)[C@H](O[C@H]4O[C@H](COP(=O)(O)OCCN)[C@@H](O)[C@H](O[C@H]5O[C@H](COP(=O)(O)OCCN)[C@@H](O)[C@H](O[C@H]6O[C@H](CO)[C@@H](O)[C@H](O[C@H]7O[C@H](CO)[C@@H](O[C@H]8O[C@H](CO[C@H]9O[C@H](CO[C@H]%10O[C@H](CO[C@H]%11O[C@H](CO[C@H]%12O[C@H](CO[C@H]%13O[C@H](CO[C@H]%14O[C@H](CO[C@H]%15O[C@H](CO[C@H]%16O[C@H](CO[C@H]%17O[C@H](CO[C@H]%18O[C@H](CO[C@H]%19O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]%19O[C@H]%19O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]%19O[C@H]%19O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]%19O)[C@@H](O)[C@H](O)[C@@H]%18O[C@H]%18O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]%18O)[C@@H](O)[C@H](O)[C@@H]%17OP(=O)(O)O[C@H]%17O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]%17O[C@H]%17O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]%17O)[C@@H](O)[C@H](O)[C@@H]%16O[C@H]%16O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]%16O[C@H]%16O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]%16O)[C@@H](O)[C@H](O)[C@@H]%15O[C@H]%15O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]%15O)[C@@H](O)[C@H](O)[C@@H]%14OP(=O)(O)O[C@H]%14O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]%14O[C@H]%14O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]%14O)[C@@H](O)[C@H](O)[C@@H]%13O[C@H]%13O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]%13O[C@H]%13O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]%13O)[C@@H](O)[C@H](O)[C@@H]%12O[C@H]%12O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]%12O)[C@@H](O)[C@H](O)[C@@H]%11OP(=O)(O)O[C@H]%11O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]%11O[C@H]%11O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]%11O)[C@@H](O)[C@H](O)[C@@H]%10O[C@H]%10O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]%10O[C@H]%10O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]%10O)[C@@H](O)[C@H](O)[C@@H]9O[C@H]9O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]9O)[C@@H](O)[C@H](O)[C@@H]8OP(=O)(O)O[C@H]8O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]8O[C@H]8O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]8O)[C@H](O[C@H]8O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]8O)[C@@H]7O)[C@@H]6O)[C@@H]5O)[C@@H]4O)[C@@H]3O)[C@@H]([C@H](O)CO)O2)[C@@H](O)[C@@H]1OC(=O)C[C@H](O)CCCCCCCCCC(C)C

SVG MW SMILES 3D mol Ionize

Warning = sub-repeat ending at aDManp: variable unit count (1-2) was assumed to be 1
Warning = sub-repeat ending at aDManp: variable unit count (1-3) was assumed to be 1
Warning = sub-repeat ending at aDManp: unknown unit count was assumed to be 3