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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...

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Autores principales: Senior, Michael J. T., Monico, Carina, Weatherill, Eve E., Gilbert, Robert J., Heuck, Alejandro P., Wallace, Mark I.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2022
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.
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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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