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Directional porin binding of intrinsically disordered protein sequences promotes colicin epitope display in the bacterial periplasm

Directional porin binding of intrinsically disordered protein sequences promotes colicin epitope display in the bacterial periplasm
Directional porin binding of intrinsically disordered protein sequences promotes colicin epitope display in the bacterial periplasm
Protein bacteriocins are potent narrow spectrum antibiotics that exploit outer membrane porins to kill bacteria by poorly understood mechanisms. Here, we determine how colicins, bacteriocins specific for Escherichia coli, engage the trimeric porin OmpF to initiate toxin entry. The N-terminal ~80 residues of the nuclease colicin ColE9 are intrinsically unstructured and house two OmpF binding sites (OBS1 and OBS2) that reside within the pores of OmpF and which flank an epitope that binds periplasmic TolB. Using a combination of molecular dynamics simulations, chemical trimerization, isothermal titration calorimetry, fluorescence microscopy and single channel recording planar lipid bilayer measurements, we show that this arrangement is achieved by OBS2 binding from the extracellular face of OmpF, whilst the interaction of OBS1 occurs from the periplasmic face of OmpF. Our study shows how the narrow pores of oligomeric porins are exploited by colicin disordered regions for direction-specific binding, which ensures the constrained presentation of an activating signal within the bacterial periplasm.
bacteria, outer membrane, colicin, intrinsic disorder, porin, OmpF, tridentate ligand
0006-2960
4374-4381
Housden, Nicholas G.
d2ad0930-bb93-4e13-a967-4ecdc84a48e4
Rassam, Patrice
2516d2de-abe9-4b88-b69a-7fa5bba29787
Lee, Sejeong
7b8ab165-fc9d-4ec0-ba26-deab261d3f74
Samsudin, Mohd
b01e87a0-af50-44d6-bca4-f511c40165f9
Kaminska, Renata
b98fd406-e88c-475d-a573-d6b2810c86d9
Sharp, Connor
2427084c-c753-446e-8c33-35f1bad53899
Goult, Jonathan D.
629a0984-6581-43e8-bbf7-dc5db6a4edf6
Francis, Marie-Louise
12b52c30-189d-4dd3-bcb0-f1e555d6c281
Khalid, Syma
90fbd954-7248-4f47-9525-4d6af9636394
Bayley, Hayley
9d13d51a-4a3d-4efe-b1b1-06e3564e23ed
Kleanthous, Colin
3d6d354c-bddf-4b64-8e4a-3f5b523802c2
Housden, Nicholas G.
d2ad0930-bb93-4e13-a967-4ecdc84a48e4
Rassam, Patrice
2516d2de-abe9-4b88-b69a-7fa5bba29787
Lee, Sejeong
7b8ab165-fc9d-4ec0-ba26-deab261d3f74
Samsudin, Mohd
b01e87a0-af50-44d6-bca4-f511c40165f9
Kaminska, Renata
b98fd406-e88c-475d-a573-d6b2810c86d9
Sharp, Connor
2427084c-c753-446e-8c33-35f1bad53899
Goult, Jonathan D.
629a0984-6581-43e8-bbf7-dc5db6a4edf6
Francis, Marie-Louise
12b52c30-189d-4dd3-bcb0-f1e555d6c281
Khalid, Syma
90fbd954-7248-4f47-9525-4d6af9636394
Bayley, Hayley
9d13d51a-4a3d-4efe-b1b1-06e3564e23ed
Kleanthous, Colin
3d6d354c-bddf-4b64-8e4a-3f5b523802c2

Housden, Nicholas G., Rassam, Patrice, Lee, Sejeong, Samsudin, Mohd, Kaminska, Renata, Sharp, Connor, Goult, Jonathan D., Francis, Marie-Louise, Khalid, Syma, Bayley, Hayley and Kleanthous, Colin (2018) Directional porin binding of intrinsically disordered protein sequences promotes colicin epitope display in the bacterial periplasm. Biochemistry, 57 (29), 4374-4381. (doi:10.1021/acs.biochem.8b00621).

Record type: Article

Abstract

Protein bacteriocins are potent narrow spectrum antibiotics that exploit outer membrane porins to kill bacteria by poorly understood mechanisms. Here, we determine how colicins, bacteriocins specific for Escherichia coli, engage the trimeric porin OmpF to initiate toxin entry. The N-terminal ~80 residues of the nuclease colicin ColE9 are intrinsically unstructured and house two OmpF binding sites (OBS1 and OBS2) that reside within the pores of OmpF and which flank an epitope that binds periplasmic TolB. Using a combination of molecular dynamics simulations, chemical trimerization, isothermal titration calorimetry, fluorescence microscopy and single channel recording planar lipid bilayer measurements, we show that this arrangement is achieved by OBS2 binding from the extracellular face of OmpF, whilst the interaction of OBS1 occurs from the periplasmic face of OmpF. Our study shows how the narrow pores of oligomeric porins are exploited by colicin disordered regions for direction-specific binding, which ensures the constrained presentation of an activating signal within the bacterial periplasm.

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Directional porin binding of intrinsically disordered protein sequences promotes colicin epitope display in the bacterial periplasm - Accepted Manuscript
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Accepted/In Press date: 27 June 2018
e-pub ahead of print date: 27 June 2018
Keywords: bacteria, outer membrane, colicin, intrinsic disorder, porin, OmpF, tridentate ligand

Identifiers

Local EPrints ID: 422185
URI: http://eprints.soton.ac.uk/id/eprint/422185
ISSN: 0006-2960
PURE UUID: a41c4d11-0cc4-4cd0-9f3b-edb2d79b2de2
ORCID for Mohd Samsudin: ORCID iD orcid.org/0000-0003-2766-4459
ORCID for Syma Khalid: ORCID iD orcid.org/0000-0002-3694-5044

Catalogue record

Date deposited: 18 Jul 2018 16:30
Last modified: 22 Nov 2021 06:32

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Contributors

Author: Nicholas G. Housden
Author: Patrice Rassam
Author: Sejeong Lee
Author: Mohd Samsudin ORCID iD
Author: Renata Kaminska
Author: Connor Sharp
Author: Jonathan D. Goult
Author: Marie-Louise Francis
Author: Syma Khalid ORCID iD
Author: Hayley Bayley
Author: Colin Kleanthous

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