Assembly of streptolysin O pores assessed by quartz crystal microbalance and atomic force microscopy provides evidence for the formation of anchored but incomplete oligomers

Sarah E Stewart, Michael E D'Angelo, Stefania Piantavigna, Rico F Tabor, Lisandra L Martin, Phillip I Bird

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Abstract

Streptolysin O (SLO) is a bacterial pore forming protein that is part of the cholesterol dependent cytolysin (CDC) family. We have used quartz crystal microbalance with dissipation monitoring (QCM-D) to examine SLO membrane binding and pore formation. In this system, SLO binds tightly to cholesterol-containing membranes, and assembles into partial and complete pores confirmed by atomic force microscopy. SLO binds to the lipid bilayer at a single rate consistent with the Langmuir isotherm model of adsorption. Changes in dissipation illustrate that SLO alters the viscoelastic properties of the bilayer during pore formation, but there is no loss of material from the bilayer as reported for small membrane-penetrating peptides. SLO mutants were used to further dissect the assembly and insertion processes by QCM-D. This shows the signature of SLO in QCM-D changes when pore formation is inhibited, and that bound and inserted SLO forms can be distinguished. Furthermore a pre-pore locked SLO mutant binds reversibly to lipid, suggesting that the partially complete wt SLO forms observed by AFM are anchored to the membrane.
Original languageEnglish
Pages (from-to)115 - 126
Number of pages12
JournalBiochimica et Biophysica Acta - Biomembranes
Volume1848
Issue number1 (Part A)
DOIs
Publication statusPublished - 2015

Cite this

@article{e38164e93235436d944815edc79e3d21,
title = "Assembly of streptolysin O pores assessed by quartz crystal microbalance and atomic force microscopy provides evidence for the formation of anchored but incomplete oligomers",
abstract = "Streptolysin O (SLO) is a bacterial pore forming protein that is part of the cholesterol dependent cytolysin (CDC) family. We have used quartz crystal microbalance with dissipation monitoring (QCM-D) to examine SLO membrane binding and pore formation. In this system, SLO binds tightly to cholesterol-containing membranes, and assembles into partial and complete pores confirmed by atomic force microscopy. SLO binds to the lipid bilayer at a single rate consistent with the Langmuir isotherm model of adsorption. Changes in dissipation illustrate that SLO alters the viscoelastic properties of the bilayer during pore formation, but there is no loss of material from the bilayer as reported for small membrane-penetrating peptides. SLO mutants were used to further dissect the assembly and insertion processes by QCM-D. This shows the signature of SLO in QCM-D changes when pore formation is inhibited, and that bound and inserted SLO forms can be distinguished. Furthermore a pre-pore locked SLO mutant binds reversibly to lipid, suggesting that the partially complete wt SLO forms observed by AFM are anchored to the membrane.",
author = "Stewart, {Sarah E} and D'Angelo, {Michael E} and Stefania Piantavigna and Tabor, {Rico F} and Martin, {Lisandra L} and Bird, {Phillip I}",
year = "2015",
doi = "10.1016/j.bbamem.2014.10.012",
language = "English",
volume = "1848",
pages = "115 -- 126",
journal = "Biochimica et Biophysica Acta - Biomembranes",
issn = "0005-2736",
publisher = "Elsevier",
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Assembly of streptolysin O pores assessed by quartz crystal microbalance and atomic force microscopy provides evidence for the formation of anchored but incomplete oligomers. / Stewart, Sarah E; D'Angelo, Michael E; Piantavigna, Stefania; Tabor, Rico F; Martin, Lisandra L; Bird, Phillip I.

In: Biochimica et Biophysica Acta - Biomembranes, Vol. 1848, No. 1 (Part A), 2015, p. 115 - 126.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Assembly of streptolysin O pores assessed by quartz crystal microbalance and atomic force microscopy provides evidence for the formation of anchored but incomplete oligomers

AU - Stewart, Sarah E

AU - D'Angelo, Michael E

AU - Piantavigna, Stefania

AU - Tabor, Rico F

AU - Martin, Lisandra L

AU - Bird, Phillip I

PY - 2015

Y1 - 2015

N2 - Streptolysin O (SLO) is a bacterial pore forming protein that is part of the cholesterol dependent cytolysin (CDC) family. We have used quartz crystal microbalance with dissipation monitoring (QCM-D) to examine SLO membrane binding and pore formation. In this system, SLO binds tightly to cholesterol-containing membranes, and assembles into partial and complete pores confirmed by atomic force microscopy. SLO binds to the lipid bilayer at a single rate consistent with the Langmuir isotherm model of adsorption. Changes in dissipation illustrate that SLO alters the viscoelastic properties of the bilayer during pore formation, but there is no loss of material from the bilayer as reported for small membrane-penetrating peptides. SLO mutants were used to further dissect the assembly and insertion processes by QCM-D. This shows the signature of SLO in QCM-D changes when pore formation is inhibited, and that bound and inserted SLO forms can be distinguished. Furthermore a pre-pore locked SLO mutant binds reversibly to lipid, suggesting that the partially complete wt SLO forms observed by AFM are anchored to the membrane.

AB - Streptolysin O (SLO) is a bacterial pore forming protein that is part of the cholesterol dependent cytolysin (CDC) family. We have used quartz crystal microbalance with dissipation monitoring (QCM-D) to examine SLO membrane binding and pore formation. In this system, SLO binds tightly to cholesterol-containing membranes, and assembles into partial and complete pores confirmed by atomic force microscopy. SLO binds to the lipid bilayer at a single rate consistent with the Langmuir isotherm model of adsorption. Changes in dissipation illustrate that SLO alters the viscoelastic properties of the bilayer during pore formation, but there is no loss of material from the bilayer as reported for small membrane-penetrating peptides. SLO mutants were used to further dissect the assembly and insertion processes by QCM-D. This shows the signature of SLO in QCM-D changes when pore formation is inhibited, and that bound and inserted SLO forms can be distinguished. Furthermore a pre-pore locked SLO mutant binds reversibly to lipid, suggesting that the partially complete wt SLO forms observed by AFM are anchored to the membrane.

UR - http://www.sciencedirect.com/science/article/pii/S0005273614003472

U2 - 10.1016/j.bbamem.2014.10.012

DO - 10.1016/j.bbamem.2014.10.012

M3 - Article

VL - 1848

SP - 115

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JO - Biochimica et Biophysica Acta - Biomembranes

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SN - 0005-2736

IS - 1 (Part A)

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