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Single‐molecule tracking of perfringolysin O assembly and membrane insertion uncoupling
We exploit single‐molecule tracking and optical single channel recording in droplet interface bilayers to resolve the assembly pathway and pore formation of the archetypical cholesterol‐dependent cytolysin nanopore, Perfringolysin O. We follow the stoichiometry and diffusion of Perfringolysin O comp...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10086847/ https://www.ncbi.nlm.nih.gov/pubmed/35989549 http://dx.doi.org/10.1111/febs.16596 |
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author | Senior, Michael J. T. Monico, Carina Weatherill, Eve E. Gilbert, Robert J. Heuck, Alejandro P. Wallace, Mark I. |
author_facet | Senior, Michael J. T. Monico, Carina Weatherill, Eve E. Gilbert, Robert J. Heuck, Alejandro P. Wallace, Mark I. |
author_sort | Senior, Michael J. T. |
collection | PubMed |
description | We exploit single‐molecule tracking and optical single channel recording in droplet interface bilayers to resolve the assembly pathway and pore formation of the archetypical cholesterol‐dependent cytolysin nanopore, Perfringolysin O. We follow the stoichiometry and diffusion of Perfringolysin O complexes during assembly with 60 ms temporal resolution and 20 nm spatial precision. Our results suggest individual nascent complexes can insert into the lipid membrane where they continue active assembly. Overall, these data support a model of stepwise irreversible assembly dominated by monomer addition, but with infrequent assembly from larger partial complexes. |
format | Online Article Text |
id | pubmed-10086847 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-100868472023-04-12 Single‐molecule tracking of perfringolysin O assembly and membrane insertion uncoupling Senior, Michael J. T. Monico, Carina Weatherill, Eve E. Gilbert, Robert J. Heuck, Alejandro P. Wallace, Mark I. FEBS J Original Articles We exploit single‐molecule tracking and optical single channel recording in droplet interface bilayers to resolve the assembly pathway and pore formation of the archetypical cholesterol‐dependent cytolysin nanopore, Perfringolysin O. We follow the stoichiometry and diffusion of Perfringolysin O complexes during assembly with 60 ms temporal resolution and 20 nm spatial precision. Our results suggest individual nascent complexes can insert into the lipid membrane where they continue active assembly. Overall, these data support a model of stepwise irreversible assembly dominated by monomer addition, but with infrequent assembly from larger partial complexes. John Wiley and Sons Inc. 2022-09-19 2023-01 /pmc/articles/PMC10086847/ /pubmed/35989549 http://dx.doi.org/10.1111/febs.16596 Text en © 2022 The Authors. The FEBS Journal published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies. https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Articles Senior, Michael J. T. Monico, Carina Weatherill, Eve E. Gilbert, Robert J. Heuck, Alejandro P. Wallace, Mark I. Single‐molecule tracking of perfringolysin O assembly and membrane insertion uncoupling |
title | Single‐molecule tracking of perfringolysin O assembly and membrane insertion uncoupling |
title_full | Single‐molecule tracking of perfringolysin O assembly and membrane insertion uncoupling |
title_fullStr | Single‐molecule tracking of perfringolysin O assembly and membrane insertion uncoupling |
title_full_unstemmed | Single‐molecule tracking of perfringolysin O assembly and membrane insertion uncoupling |
title_short | Single‐molecule tracking of perfringolysin O assembly and membrane insertion uncoupling |
title_sort | single‐molecule tracking of perfringolysin o assembly and membrane insertion uncoupling |
topic | Original Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10086847/ https://www.ncbi.nlm.nih.gov/pubmed/35989549 http://dx.doi.org/10.1111/febs.16596 |
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