Colicin E1 opens its hinge to plug TolC
The double membrane architecture of Gram-negative bacteria forms a barrier that is impermeable to most extracellular threats. Bacteriocin proteins evolved to exploit the accessible, surface-exposed proteins embedded in the outer membrane to deliver cytotoxic cargo. Colicin E1 is a bacteriocin produc...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2022-02-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/73297 |
| _version_ | 1828169827803463680 |
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| author | S Jimmy Budiardjo Jacqueline J Stevens Anna L Calkins Ayotunde P Ikujuni Virangika K Wimalasena Emre Firlar David A Case Julie S Biteen Jason T Kaelber Joanna SG Slusky |
| author_facet | S Jimmy Budiardjo Jacqueline J Stevens Anna L Calkins Ayotunde P Ikujuni Virangika K Wimalasena Emre Firlar David A Case Julie S Biteen Jason T Kaelber Joanna SG Slusky |
| author_sort | S Jimmy Budiardjo |
| collection | DOAJ |
| description | The double membrane architecture of Gram-negative bacteria forms a barrier that is impermeable to most extracellular threats. Bacteriocin proteins evolved to exploit the accessible, surface-exposed proteins embedded in the outer membrane to deliver cytotoxic cargo. Colicin E1 is a bacteriocin produced by, and lethal to, Escherichia coli that hijacks the outer membrane proteins (OMPs) TolC and BtuB to enter the cell. Here, we capture the colicin E1 translocation domain inside its membrane receptor, TolC, by high-resolution cryo-electron microscopy to obtain the first reported structure of a bacteriocin bound to TolC. Colicin E1 binds stably to TolC as an open hinge through the TolC pore—an architectural rearrangement from colicin E1’s unbound conformation. This binding is stable in live E. coli cells as indicated by single-molecule fluorescence microscopy. Finally, colicin E1 fragments binding to TolC plug the channel, inhibiting its native efflux function as an antibiotic efflux pump, and heightening susceptibility to three antibiotic classes. In addition to demonstrating that these protein fragments are useful starting points for developing novel antibiotic potentiators, this method could be expanded to other colicins to inhibit other OMP functions. |
| first_indexed | 2024-04-12T02:58:45Z |
| format | Article |
| id | doaj.art-9510bb93037e456488cca7fe070b1f34 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T02:58:45Z |
| publishDate | 2022-02-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-9510bb93037e456488cca7fe070b1f342022-12-22T03:50:43ZengeLife Sciences Publications LtdeLife2050-084X2022-02-011110.7554/eLife.73297Colicin E1 opens its hinge to plug TolCS Jimmy Budiardjo0https://orcid.org/0000-0003-2094-9179Jacqueline J Stevens1https://orcid.org/0000-0003-2235-0522Anna L Calkins2Ayotunde P Ikujuni3https://orcid.org/0000-0001-8951-3440Virangika K Wimalasena4https://orcid.org/0000-0002-1061-3439Emre Firlar5https://orcid.org/0000-0003-0190-6528David A Case6Julie S Biteen7https://orcid.org/0000-0003-2038-6484Jason T Kaelber8https://orcid.org/0000-0001-9426-1030Joanna SG Slusky9https://orcid.org/0000-0003-0842-6340Center for Computational Biology, The University of Kansas, Lawrence, United StatesDepartment of Molecular Biosciences, The University of Kansas, Lawrence, United StatesDepartment of Chemistry, University of Michigan, Ann Arbor, United StatesDepartment of Molecular Biosciences, The University of Kansas, Lawrence, United StatesDepartment of Molecular Biosciences, The University of Kansas, Lawrence, United StatesRutgers CryoEM & Nanoimaging Facility and Institute for Quantitative Biomedicine, Rutgers University, Piscataway, United StatesDepartment of Chemistry and Chemical Biology, Rutgers University, Piscataway, United StatesDepartment of Chemistry, University of Michigan, Ann Arbor, United StatesRutgers CryoEM & Nanoimaging Facility and Institute for Quantitative Biomedicine, Rutgers University, Piscataway, United StatesCenter for Computational Biology, The University of Kansas, Lawrence, United States; Department of Molecular Biosciences, The University of Kansas, Lawrence, United StatesThe double membrane architecture of Gram-negative bacteria forms a barrier that is impermeable to most extracellular threats. Bacteriocin proteins evolved to exploit the accessible, surface-exposed proteins embedded in the outer membrane to deliver cytotoxic cargo. Colicin E1 is a bacteriocin produced by, and lethal to, Escherichia coli that hijacks the outer membrane proteins (OMPs) TolC and BtuB to enter the cell. Here, we capture the colicin E1 translocation domain inside its membrane receptor, TolC, by high-resolution cryo-electron microscopy to obtain the first reported structure of a bacteriocin bound to TolC. Colicin E1 binds stably to TolC as an open hinge through the TolC pore—an architectural rearrangement from colicin E1’s unbound conformation. This binding is stable in live E. coli cells as indicated by single-molecule fluorescence microscopy. Finally, colicin E1 fragments binding to TolC plug the channel, inhibiting its native efflux function as an antibiotic efflux pump, and heightening susceptibility to three antibiotic classes. In addition to demonstrating that these protein fragments are useful starting points for developing novel antibiotic potentiators, this method could be expanded to other colicins to inhibit other OMP functions.https://elifesciences.org/articles/73297antibiotic effluxcolicinantibiotic resistanceTolCcolicin E1 |
| spellingShingle | S Jimmy Budiardjo Jacqueline J Stevens Anna L Calkins Ayotunde P Ikujuni Virangika K Wimalasena Emre Firlar David A Case Julie S Biteen Jason T Kaelber Joanna SG Slusky Colicin E1 opens its hinge to plug TolC eLife antibiotic efflux colicin antibiotic resistance TolC colicin E1 |
| title | Colicin E1 opens its hinge to plug TolC |
| title_full | Colicin E1 opens its hinge to plug TolC |
| title_fullStr | Colicin E1 opens its hinge to plug TolC |
| title_full_unstemmed | Colicin E1 opens its hinge to plug TolC |
| title_short | Colicin E1 opens its hinge to plug TolC |
| title_sort | colicin e1 opens its hinge to plug tolc |
| topic | antibiotic efflux colicin antibiotic resistance TolC colicin E1 |
| url | https://elifesciences.org/articles/73297 |
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