Peptidoglycan binding by a pocket on the accessory NTF2-domain of Pgp2 directs helical cell shape of Campylobacter jejuni
| Autor: | Martin E. Tanner, Jacob A. Brockerman, Chang Sheng-Huei Lin, Jean-Pierre Simorre, Erin C. Gaynor, Jenny Vermeulen, Lawrence P. McIntosh, Anson C. K. Chan, Arvind Soni, Michael E. P. Murphy |
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| Přispěvatelé: | Department of Microbiology and Immunology [Vancouver] (UBC Microbiology), University of British Columbia (UBC), Institut de biologie structurale (IBS - UMR 5075), Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA) |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Nucleocytoplasmic Transport Proteins Protein Conformation bacterial cell shape Mutant MESH: Catalytic Domain Peptide Carboxypeptidases peptidoglycan Biochemistry chemistry.chemical_compound MESH: Protein Conformation Catalytic Domain MESH: Bacterial Proteins chemistry.chemical_classification biology NTF2 domain [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry Molecular Biology/Structural Biology [q-bio.BM] MESH: Nucleocytoplasmic Transport Proteins MESH: Peptidoglycan PG peptidoglycan carboxypeptidase HPLC high performance liquid chromatography docking Peptidoglycan binding Research Article crystal structure nuclear magnetic resonance (NMR) Campylobacter jejuni MESH: Campylobacter jejuni 03 medical and health sciences Bacterial Proteins Humans Molecular Biology MESH: Humans 030102 biochemistry & molecular biology Active site Campylobacter Cell Biology Periplasmic space biology.organism_classification AIR ambiguous interaction restraint CSP chemical shift perturbation Carboxypeptidase 030104 developmental biology chemistry biology.protein Biophysics Peptidoglycan MESH: Carboxypeptidases |
| Zdroj: | Journal of Biological Chemistry Journal of Biological Chemistry, 2021, 296, pp.100528. ⟨10.1016/j.jbc.2021.100528⟩ Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2021, 296, pp.100528. ⟨10.1016/j.jbc.2021.100528⟩ The Journal of Biological Chemistry |
| ISSN: | 0021-9258 1083-351X |
| DOI: | 10.1016/j.jbc.2021.100528⟩ |
| Popis: | International audience; The helical morphology of Campylobacter jejuni, a bacterium involved in host gut colonization and pathogenesis in humans, is determined by the structure of the peptidoglycan (PG) layer. This structure is dictated by trimming of peptide stems by the LD-carboxypeptidase Pgp2 within the periplasm. The interaction interface between Pgp2 and PG to select sites for peptide trimming is unknown. We determined a 1.6 Å resolution crystal structure of Pgp2, which contains a conserved LD-carboxypeptidase domain and a previously uncharacterized domain with an NTF2-like fold (NTF2). We identified a pocket in the NTF2 domain formed by conserved residues and located ∼40 Å from the LD-carboxypeptidase active site. Expression of pgp2 in trans with substitutions of charged (Lys257, Lys307, Glu324) and hydrophobic residues (Phe242 and Tyr233) within the pocket did not restore helical morphology to a pgp2 deletion strain. Muropeptide analysis indicated a decrease of murotripeptides in the deletion strain expressing these mutants, suggesting reduced Pgp2 catalytic activity. Pgp2 but not the K307A mutant was pulled down by C. jejuni Δpgp2 PG sacculi, supporting a role for the pocket in PG binding. NMR spectroscopy was used to define the interaction interfaces of Pgp2 with several PG fragments, which bound to the active site within the LD-carboxypeptidase domain and the pocket of the NTF2 domain. We propose a model for Pgp2 binding to PG strands involving both the LD-carboxypeptidase domain and the accessory NTF2 domain to induce a helical cell shape. |
| Databáze: | OpenAIRE |
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