The Structural Basis for a Transition State That Regulates Pore Formation in a Bacterial Toxin

ABSTRACT The cholesterol-dependent cytolysin (CDC) genes are present in bacterial species that span terrestrial, vertebrate, and invertebrate niches, which suggests that they have evolved to function under widely different environmental conditions. Using a combination of biophysical and crystallogra...

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Autores principales:	Kristin R. Wade, Sara L. Lawrence, Allison J. Farrand, Eileen M. Hotze, Michael J. Kuiper, Michael A. Gorman, Michelle P. Christie, Santosh Panjikar, Craig J. Morton, Michael W. Parker, Rodney K. Tweten
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	cold denaturation complement gasdermin perforin water network Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/7bf35b5ddc6847bfb07b9a7eda692855
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spelling	oai:doaj.org-article:7bf35b5ddc6847bfb07b9a7eda6928552021-11-15T15:55:26ZThe Structural Basis for a Transition State That Regulates Pore Formation in a Bacterial Toxin10.1128/mBio.00538-192150-7511https://doaj.org/article/7bf35b5ddc6847bfb07b9a7eda6928552019-04-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00538-19https://doaj.org/toc/2150-7511ABSTRACT The cholesterol-dependent cytolysin (CDC) genes are present in bacterial species that span terrestrial, vertebrate, and invertebrate niches, which suggests that they have evolved to function under widely different environmental conditions. Using a combination of biophysical and crystallographic approaches, we reveal that the relative stability of an intramolecular interface in the archetype CDC perfringolysin O (PFO) plays a central role in regulating its pore-forming properties. The disruption of this interface allows the formation of the membrane spanning β-barrel pore in all CDCs. We show here that the relative strength of the stabilizing forces at this interface directly impacts the energy barrier posed by the transition state for pore formation, as reflected in the Arrhenius activation energy (Ea) for pore formation. This change directly impacts the kinetics and temperature dependence of pore formation. We further show that the interface structure in a CDC from a terrestrial species enables it to function efficiently across a wide range of temperatures by minimizing changes in the strength of the transition state barrier to pore formation. These studies establish a paradigm that CDCs, and possibly other β-barrel pore-forming proteins/toxins, can evolve significantly different pore-forming properties by altering the stability of this transitional interface, which impacts the kinetic parameters and temperature dependence of pore formation. IMPORTANCE The cholesterol-dependent cytolysins (CDCs) are the archetype for the superfamily of oligomeric pore-forming proteins that includes the membrane attack complex/perforin (MACPF) family of immune defense proteins and the stonefish venom toxins (SNTX). The CDC/MACPF/SNTX family exhibits a common protein fold, which forms a membrane-spanning β-barrel pore. We show that changing the relative stability of an extensive intramolecular interface within this fold, which is necessarily disrupted to form the large β-barrel pore, dramatically alters the kinetic and temperature-dependent properties of CDC pore formation. These studies show that the CDCs and other members of the CDC/MACPF/SNTX superfamily have the capacity to significantly alter their pore-forming properties to function under widely different environmental conditions encountered by these species.Kristin R. WadeSara L. LawrenceAllison J. FarrandEileen M. HotzeMichael J. KuiperMichael A. GormanMichelle P. ChristieSantosh PanjikarCraig J. MortonMichael W. ParkerRodney K. TwetenAmerican Society for Microbiologyarticlecold denaturationcomplementgasderminperforinwater networkMicrobiologyQR1-502ENmBio, Vol 10, Iss 2 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	cold denaturation complement gasdermin perforin water network Microbiology QR1-502
spellingShingle	cold denaturation complement gasdermin perforin water network Microbiology QR1-502 Kristin R. Wade Sara L. Lawrence Allison J. Farrand Eileen M. Hotze Michael J. Kuiper Michael A. Gorman Michelle P. Christie Santosh Panjikar Craig J. Morton Michael W. Parker Rodney K. Tweten The Structural Basis for a Transition State That Regulates Pore Formation in a Bacterial Toxin
description	ABSTRACT The cholesterol-dependent cytolysin (CDC) genes are present in bacterial species that span terrestrial, vertebrate, and invertebrate niches, which suggests that they have evolved to function under widely different environmental conditions. Using a combination of biophysical and crystallographic approaches, we reveal that the relative stability of an intramolecular interface in the archetype CDC perfringolysin O (PFO) plays a central role in regulating its pore-forming properties. The disruption of this interface allows the formation of the membrane spanning β-barrel pore in all CDCs. We show here that the relative strength of the stabilizing forces at this interface directly impacts the energy barrier posed by the transition state for pore formation, as reflected in the Arrhenius activation energy (Ea) for pore formation. This change directly impacts the kinetics and temperature dependence of pore formation. We further show that the interface structure in a CDC from a terrestrial species enables it to function efficiently across a wide range of temperatures by minimizing changes in the strength of the transition state barrier to pore formation. These studies establish a paradigm that CDCs, and possibly other β-barrel pore-forming proteins/toxins, can evolve significantly different pore-forming properties by altering the stability of this transitional interface, which impacts the kinetic parameters and temperature dependence of pore formation. IMPORTANCE The cholesterol-dependent cytolysins (CDCs) are the archetype for the superfamily of oligomeric pore-forming proteins that includes the membrane attack complex/perforin (MACPF) family of immune defense proteins and the stonefish venom toxins (SNTX). The CDC/MACPF/SNTX family exhibits a common protein fold, which forms a membrane-spanning β-barrel pore. We show that changing the relative stability of an extensive intramolecular interface within this fold, which is necessarily disrupted to form the large β-barrel pore, dramatically alters the kinetic and temperature-dependent properties of CDC pore formation. These studies show that the CDCs and other members of the CDC/MACPF/SNTX superfamily have the capacity to significantly alter their pore-forming properties to function under widely different environmental conditions encountered by these species.
format	article
author	Kristin R. Wade Sara L. Lawrence Allison J. Farrand Eileen M. Hotze Michael J. Kuiper Michael A. Gorman Michelle P. Christie Santosh Panjikar Craig J. Morton Michael W. Parker Rodney K. Tweten
author_facet	Kristin R. Wade Sara L. Lawrence Allison J. Farrand Eileen M. Hotze Michael J. Kuiper Michael A. Gorman Michelle P. Christie Santosh Panjikar Craig J. Morton Michael W. Parker Rodney K. Tweten
author_sort	Kristin R. Wade
title	The Structural Basis for a Transition State That Regulates Pore Formation in a Bacterial Toxin
title_short	The Structural Basis for a Transition State That Regulates Pore Formation in a Bacterial Toxin
title_full	The Structural Basis for a Transition State That Regulates Pore Formation in a Bacterial Toxin
title_fullStr	The Structural Basis for a Transition State That Regulates Pore Formation in a Bacterial Toxin
title_full_unstemmed	The Structural Basis for a Transition State That Regulates Pore Formation in a Bacterial Toxin
title_sort	structural basis for a transition state that regulates pore formation in a bacterial toxin
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/7bf35b5ddc6847bfb07b9a7eda692855
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The Structural Basis for a Transition State That Regulates Pore Formation in a Bacterial Toxin

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