Skip to main content
bioRxiv
  • Home
  • About
  • Submit
  • ALERTS / RSS
Advanced Search
New Results

Single molecule tracking the uncoupling of assembly and membrane insertion in Perfringolysin O

Michael J T Senior, Carina Monico, Eve E Weatherill, Robert J Gilbert, Alejandro P Heuck, View ORCID ProfileMark I Wallace
doi: https://doi.org/10.1101/2021.05.26.445776
Michael J T Senior
†Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford OX1 3TA, United Kingdom
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Carina Monico
‡Department of Chemistry, King’s College London, London SE1 1DB, United Kingdom
†Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford OX1 3TA, United Kingdom
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Eve E Weatherill
‡Department of Chemistry, King’s College London, London SE1 1DB, United Kingdom
†Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford OX1 3TA, United Kingdom
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Robert J Gilbert
¶Division of Structural Biology, Wellcome Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, United Kingdom
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Alejandro P Heuck
§Departments of Biochemistry and Molecular Biology, University of Massachusetts, Amherst, MA, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Mark I Wallace
‡Department of Chemistry, King’s College London, London SE1 1DB, United Kingdom
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Mark I Wallace
  • For correspondence: mark.wallace@kcl.ac.uk
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Supplementary material
  • Preview PDF
Loading

Abstract

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 millisecond temporal resolution and 20 nanometre 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.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • Changes to format and discussion.

  • Abbreviations

    CDC
    (Cholesterol-Dependant Cytolysin)
    PFO
    (Perfringolysin O)
    MACPF
    (Membrane Attack Complex)
    DIB
    (Droplet Interface Bilayer)
    AFM
    (Atomic Force Mi-croscopy)
    TIRF
    (Total Internal Reflection)
    oSCR
    (Optical Single-Channel Recording)
  • Copyright 
    The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC 4.0 International license.
    Back to top
    PreviousNext
    Posted June 23, 2022.
    Download PDF

    Supplementary Material

    Email

    Thank you for your interest in spreading the word about bioRxiv.

    NOTE: Your email address is requested solely to identify you as the sender of this article.

    Enter multiple addresses on separate lines or separate them with commas.
    Single molecule tracking the uncoupling of assembly and membrane insertion in Perfringolysin O
    (Your Name) has forwarded a page to you from bioRxiv
    (Your Name) thought you would like to see this page from the bioRxiv website.
    CAPTCHA
    This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
    Share
    Single molecule tracking the uncoupling of assembly and membrane insertion in Perfringolysin O
    Michael J T Senior, Carina Monico, Eve E Weatherill, Robert J Gilbert, Alejandro P Heuck, Mark I Wallace
    bioRxiv 2021.05.26.445776; doi: https://doi.org/10.1101/2021.05.26.445776
    Reddit logo Twitter logo Facebook logo LinkedIn logo Mendeley logo
    Citation Tools
    Single molecule tracking the uncoupling of assembly and membrane insertion in Perfringolysin O
    Michael J T Senior, Carina Monico, Eve E Weatherill, Robert J Gilbert, Alejandro P Heuck, Mark I Wallace
    bioRxiv 2021.05.26.445776; doi: https://doi.org/10.1101/2021.05.26.445776

    Citation Manager Formats

    • BibTeX
    • Bookends
    • EasyBib
    • EndNote (tagged)
    • EndNote 8 (xml)
    • Medlars
    • Mendeley
    • Papers
    • RefWorks Tagged
    • Ref Manager
    • RIS
    • Zotero
    • Tweet Widget
    • Facebook Like
    • Google Plus One

    Subject Area

    • Biophysics
    Subject Areas
    All Articles
    • Animal Behavior and Cognition (4238)
    • Biochemistry (9160)
    • Bioengineering (6797)
    • Bioinformatics (24054)
    • Biophysics (12149)
    • Cancer Biology (9564)
    • Cell Biology (13821)
    • Clinical Trials (138)
    • Developmental Biology (7654)
    • Ecology (11733)
    • Epidemiology (2066)
    • Evolutionary Biology (15536)
    • Genetics (10665)
    • Genomics (14353)
    • Immunology (9504)
    • Microbiology (22887)
    • Molecular Biology (9121)
    • Neuroscience (49094)
    • Paleontology (357)
    • Pathology (1487)
    • Pharmacology and Toxicology (2579)
    • Physiology (3851)
    • Plant Biology (8349)
    • Scientific Communication and Education (1473)
    • Synthetic Biology (2300)
    • Systems Biology (6204)
    • Zoology (1302)