Structural evidence that colicin a protein binds to a novel binding site of TolA protein in Escherichia coli periplasm
The Tol assembly of proteins is an interacting network of proteins located in the Escherichia coli cell envelope that transduces energy and contributes to cell integrity. TolA is central to this network linking the inner and outer membranes by interactions with TolQ, TolR, TolB, and Pal. Group A col...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
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American Society for Biochemistry and Molecular Biology
2012
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| Online Access: | https://eprints.nottingham.ac.uk/2439/ |
| _version_ | 1848790785905917952 |
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| author | Li, Chan Zhang, Ying Vankemmelbeke, Mireille Hecht, Oliver Aleanizy, Fadilah Sfouq Macdonald, Colin Moore, Geoffrey R. James, Richard Penfold, Christopher N. |
| author_facet | Li, Chan Zhang, Ying Vankemmelbeke, Mireille Hecht, Oliver Aleanizy, Fadilah Sfouq Macdonald, Colin Moore, Geoffrey R. James, Richard Penfold, Christopher N. |
| author_sort | Li, Chan |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | The Tol assembly of proteins is an interacting network of proteins located in the Escherichia coli cell envelope that transduces energy and contributes to cell integrity. TolA is central to this network linking the inner and outer membranes by interactions with TolQ, TolR, TolB, and Pal. Group A colicins, such as ColA, parasitize the Tol network through interactions with TolA and/or TolB to facilitate translocation through the cell envelope to reach their cytotoxic site of action. We have determined the first structure of the C-terminal domain of TolA (TolAIII) bound to an N-terminal ColA polypeptide (TA53–107). The interface region of the TA53–107-TolAIII complex consists of polar contacts linking residues Arg-92 to Arg-96 of ColA with residues Leu-375–Pro-380 of TolA, which constitutes a β-strand addition commonly seen in more promiscuous protein-protein contacts. The interface region also includes three cation-π interactions (Tyr-58–Lys-368, Tyr-90–Lys-379, Phe-94–Lys-396), which have not been observed in any other colicin-Tol protein complex. Mutagenesis of the interface residues of ColA or TolA revealed that the effect on the interaction was cumulative; single mutations of either partner had no effect on ColA activity, whereas mutations of three or more residues significantly reduced ColA activity. Mutagenesis of the aromatic ring component of the cation-π interacting residues showed Tyr-58 of ColA to be essential for the stability of complex formation. TA53–107 binds on the opposite side of TolAIII to that used by g3p, ColN, or TolB, illustrating the flexible nature of TolA as a periplasmic hub protein. |
| first_indexed | 2025-11-14T18:18:08Z |
| format | Article |
| id | nottingham-2439 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T18:18:08Z |
| publishDate | 2012 |
| publisher | American Society for Biochemistry and Molecular Biology |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-24392020-05-04T16:32:57Z https://eprints.nottingham.ac.uk/2439/ Structural evidence that colicin a protein binds to a novel binding site of TolA protein in Escherichia coli periplasm Li, Chan Zhang, Ying Vankemmelbeke, Mireille Hecht, Oliver Aleanizy, Fadilah Sfouq Macdonald, Colin Moore, Geoffrey R. James, Richard Penfold, Christopher N. The Tol assembly of proteins is an interacting network of proteins located in the Escherichia coli cell envelope that transduces energy and contributes to cell integrity. TolA is central to this network linking the inner and outer membranes by interactions with TolQ, TolR, TolB, and Pal. Group A colicins, such as ColA, parasitize the Tol network through interactions with TolA and/or TolB to facilitate translocation through the cell envelope to reach their cytotoxic site of action. We have determined the first structure of the C-terminal domain of TolA (TolAIII) bound to an N-terminal ColA polypeptide (TA53–107). The interface region of the TA53–107-TolAIII complex consists of polar contacts linking residues Arg-92 to Arg-96 of ColA with residues Leu-375–Pro-380 of TolA, which constitutes a β-strand addition commonly seen in more promiscuous protein-protein contacts. The interface region also includes three cation-π interactions (Tyr-58–Lys-368, Tyr-90–Lys-379, Phe-94–Lys-396), which have not been observed in any other colicin-Tol protein complex. Mutagenesis of the interface residues of ColA or TolA revealed that the effect on the interaction was cumulative; single mutations of either partner had no effect on ColA activity, whereas mutations of three or more residues significantly reduced ColA activity. Mutagenesis of the aromatic ring component of the cation-π interacting residues showed Tyr-58 of ColA to be essential for the stability of complex formation. TA53–107 binds on the opposite side of TolAIII to that used by g3p, ColN, or TolB, illustrating the flexible nature of TolA as a periplasmic hub protein. American Society for Biochemistry and Molecular Biology 2012-04-09 Article PeerReviewed Li, Chan, Zhang, Ying, Vankemmelbeke, Mireille, Hecht, Oliver, Aleanizy, Fadilah Sfouq, Macdonald, Colin, Moore, Geoffrey R., James, Richard and Penfold, Christopher N. (2012) Structural evidence that colicin a protein binds to a novel binding site of TolA protein in Escherichia coli periplasm. The Journal of Biological Chemistry, 287 (23). pp. 19048-19057. ISSN 0021-9258 Bacterial Toxins Microbiology Protein Structure Protein-Protein Interactions X-ray Crystallography Tol Bacteriocin Colicin http://www.jbc.org/content/287/23/19048.long doi:10.1074/jbc.M112.342246 doi:10.1074/jbc.M112.342246 |
| spellingShingle | Bacterial Toxins Microbiology Protein Structure Protein-Protein Interactions X-ray Crystallography Tol Bacteriocin Colicin Li, Chan Zhang, Ying Vankemmelbeke, Mireille Hecht, Oliver Aleanizy, Fadilah Sfouq Macdonald, Colin Moore, Geoffrey R. James, Richard Penfold, Christopher N. Structural evidence that colicin a protein binds to a novel binding site of TolA protein in Escherichia coli periplasm |
| title | Structural evidence that colicin a protein binds to a novel binding site of TolA protein in Escherichia coli periplasm |
| title_full | Structural evidence that colicin a protein binds to a novel binding site of TolA protein in Escherichia coli periplasm |
| title_fullStr | Structural evidence that colicin a protein binds to a novel binding site of TolA protein in Escherichia coli periplasm |
| title_full_unstemmed | Structural evidence that colicin a protein binds to a novel binding site of TolA protein in Escherichia coli periplasm |
| title_short | Structural evidence that colicin a protein binds to a novel binding site of TolA protein in Escherichia coli periplasm |
| title_sort | structural evidence that colicin a protein binds to a novel binding site of tola protein in escherichia coli periplasm |
| topic | Bacterial Toxins Microbiology Protein Structure Protein-Protein Interactions X-ray Crystallography Tol Bacteriocin Colicin |
| url | https://eprints.nottingham.ac.uk/2439/ https://eprints.nottingham.ac.uk/2439/ https://eprints.nottingham.ac.uk/2439/ |