Single-molecule tracking of perfringolysin O assembly and membrane insertion uncoupling

Michael J.T. Senior; Carina Monico; Eve E. Weatherill; Robert J. Gilbert; Alejandro P. Heuck; Mark I. Wallace

doi:10.1111/febs.16596

Single-molecule tracking of perfringolysin O assembly and membrane insertion uncoupling

Michael J.T. Senior, Carina Monico, Eve E. Weatherill, Robert J. Gilbert, Alejandro P. Heuck, Mark I. Wallace^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

3 Citations (Scopus)

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 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.

Original language	English
Journal	FEBS Journal
DOIs	https://doi.org/10.1111/febs.16596
Publication status	Accepted/In press - 2022

Keywords

assembly
DIB
fluorescence
PFO
photobleaching
single molecule

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10.1111/febs.16596

Cite this

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title = "Single-molecule tracking of perfringolysin O assembly and membrane insertion uncoupling",

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 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.",

keywords = "assembly, DIB, fluorescence, PFO, photobleaching, single molecule",

author = "Senior, {Michael J.T.} and Carina Monico and Weatherill, {Eve E.} and Gilbert, {Robert J.} and Heuck, {Alejandro P.} and Wallace, {Mark I.}",

note = "Funding Information: MJS was funded by the EPSRC Life‐Science Interface Doctoral Training Centre at Oxford University. We thank Till B{\"o}cking and James Walsh for their helpful comments during the preparation of this manuscript. Publisher Copyright: {\textcopyright} 2022 The Authors. The FEBS Journal published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.",

year = "2022",

doi = "10.1111/febs.16596",

language = "English",

journal = "FEBS Journal",

issn = "1742-464X",

publisher = "Wiley",

}

TY - JOUR

T1 - Single-molecule tracking of perfringolysin O assembly and membrane insertion uncoupling

AU - Senior, Michael J.T.

AU - Monico, Carina

AU - Weatherill, Eve E.

AU - Gilbert, Robert J.

AU - Heuck, Alejandro P.

AU - Wallace, Mark I.

N1 - Funding Information: MJS was funded by the EPSRC Life‐Science Interface Doctoral Training Centre at Oxford University. We thank Till Böcking and James Walsh for their helpful comments during the preparation of this manuscript. Publisher Copyright: © 2022 The Authors. The FEBS Journal published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.

PY - 2022

Y1 - 2022

N2 - 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.

AB - 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.

KW - assembly

KW - DIB

KW - fluorescence

KW - PFO

KW - photobleaching

KW - single molecule

UR - http://www.scopus.com/inward/record.url?scp=85138265822&partnerID=8YFLogxK

U2 - 10.1111/febs.16596

DO - 10.1111/febs.16596

M3 - Article

C2 - 35989549

AN - SCOPUS:85138265822

SN - 1742-464X

JO - FEBS Journal

JF - FEBS Journal

ER -

Single-molecule tracking of perfringolysin O assembly and membrane insertion uncoupling

Abstract

Keywords

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

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Single-molecule tracking of perfringolysin O assembly and membrane insertion uncoupling

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

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