Structure (London), 2019-07, Vol.27 (7), p.1082-1093.e5
2019
Details
- Autor(en) / Beteiligte
- Titel
- Structure and Assembly of the Enterohemorrhagic Escherichia coli Type 4 Pilus
- Ist Teil von
- Structure (London), 2019-07, Vol.27 (7), p.1082-1093.e5
- Ort / Verlag
- United States: Elsevier Ltd
- Erscheinungsjahr
- 2019
- Link zum Volltext
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- Bacterial type 4a pili are dynamic surface filaments that promote bacterial adherence, motility, and macromolecular transport. Their genes are highly conserved among enterobacteria and their expression in enterohemorrhagic Escherichia coli (EHEC) promotes adhesion to intestinal epithelia and pro-inflammatory signaling. To define the molecular basis of EHEC pilus assembly, we determined the structure of the periplasmic domain of its major subunit PpdD (PpdDp), a prototype of an enterobacterial pilin subfamily containing two disulfide bonds. The structure of PpdDp, determined by NMR, was then docked into the density envelope of purified EHEC pili obtained by cryoelectron microscopy (cryo-EM). Cryo-EM reconstruction of EHEC pili at ∼8 Å resolution revealed extremely high pilus flexibility correlating with a large extended region of the pilin stem. Systematic mutagenesis combined with functional and interaction analyses identified charged residues essential for pilus assembly. Structural information on exposed regions and interfaces between EHEC pilins is relevant for vaccine and drug discovery. [Display omitted] •EHEC Type 4a pili are the prototype of competence co-regulated enterobacterial pili•Combined NMR and cryo-EM analysis provided the structure of EHEC type 4 pili•Major pilin displays a long, melted N-terminal segment in highly flexible EHEC pili•Electrostatic contacts between pilins and assembly factors drive pilus assembly Atomic model of type 4a pili from enterohemorrhagic Escherichia coli is obtained by combining NMR structure of the major pilin PpdD and the cryo-EM map of assembled pili. Mutational analysis of PpdD identified electrostatic contacts between pilins and revealed interactions with specific assembly factors involved in pilus assembly.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 0969-2126eISSN: 1878-4186DOI: 10.1016/j.str.2019.03.021Titel-ID: cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_7003672
- Format
- –
- Schlagworte
- Amino Acid Sequence, Bacteriology, Binding Sites, Biochemistry, Molecular Biology, Cloning, Molecular, cryo-EM, Cryoelectron Microscopy, EHEC, Enterohemorrhagic Escherichia coli - chemistry, Enterohemorrhagic Escherichia coli - metabolism, Escherichia coli, Escherichia coli - genetics, Escherichia coli - metabolism, Escherichia coli Proteins - chemistry, Escherichia coli Proteins - genetics, Escherichia coli Proteins - metabolism, Fimbriae Proteins - chemistry, Fimbriae Proteins - genetics, Fimbriae Proteins - metabolism, Fimbriae, Bacterial - chemistry, Fimbriae, Bacterial - metabolism, Fimbriae, Bacterial - ultrastructure, Gene Expression, Genetic Vectors - chemistry, Genetic Vectors - metabolism, hemorrhagic coli pilus (HCP), Kinetics, Life Sciences, Microbiology and Parasitology, Molecular Docking Simulation, molecular modeling, Mutation, NMR, Nuclear Magnetic Resonance, Biomolecular, pilin structure, pilus assembly, Protein Binding, Protein Conformation, alpha-Helical, Protein Conformation, beta-Strand, Protein Interaction Domains and Motifs, Quantitative Methods, Recombinant Proteins - chemistry, Recombinant Proteins - genetics, Recombinant Proteins - metabolism, Sequence Alignment, Sequence Homology, Amino Acid, Static Electricity, Structural Biology, Thermodynamics, type 4 pili, type II secretion system