19F NMR studies provide insights into lipid membrane interactions of listeriolysin O, a pore forming toxin from Listeria monocytogenes

Abstract Listeria monocytogenes is a mammalian pathogen that causes gastroenteritis, miscarriages and infections of the central nervous system in immunocompromised individuals. Its main virulence factor is listeriolysin O (LLO), a pore-forming cholesterol-dependent cytolysin (CDC), which enables bac...

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Autores principales:	Mirijam Kozorog, Marc-Antoine Sani, Martina Lenarčič Živković, Gregor Ilc, Vesna Hodnik, Frances Separovic, Janez Plavec, Gregor Anderluh
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d6860f4192aa49c7bb6735ed309dc05f
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spelling	oai:doaj.org-article:d6860f4192aa49c7bb6735ed309dc05f2021-12-02T15:08:37Z19F NMR studies provide insights into lipid membrane interactions of listeriolysin O, a pore forming toxin from Listeria monocytogenes10.1038/s41598-018-24692-62045-2322https://doaj.org/article/d6860f4192aa49c7bb6735ed309dc05f2018-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-24692-6https://doaj.org/toc/2045-2322Abstract Listeria monocytogenes is a mammalian pathogen that causes gastroenteritis, miscarriages and infections of the central nervous system in immunocompromised individuals. Its main virulence factor is listeriolysin O (LLO), a pore-forming cholesterol-dependent cytolysin (CDC), which enables bacterial escape from the phagolysosome and contributes to bacterial pathogenicity. Details of cholesterol (Chol) recognition and membrane binding mechanisms by LLO are still not known. Here we used 19F-NMR spectroscopy in order to assess LLO-Chol interactions in solution and in a Chol-rich membrane environment. LLO has six tryptophan residues located in the region of the molecule that is first in contact with lipid membranes. 19F-LLO, which contained 5-fluoro-tryptophans, was prepared by using isotopic labelling in an E. coli expression system. Signals in the 19F-NMR spectrum of 19F-LLO were unambiguously assigned by using a series of single Trp → Phe point mutations. The results employing various cholesterol preparations in solution indicate that tryptophan residues are not directly involved in Chol binding in solution. However, significant chemical shift changes were observed upon LLO binding to Chol-rich membranes, highlighting the role of tryptophan residues in membrane interactions (W512) and oligomerisation (W189 and W489).Mirijam KozorogMarc-Antoine SaniMartina Lenarčič ŽivkovićGregor IlcVesna HodnikFrances SeparovicJanez PlavecGregor AnderluhNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-11 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Mirijam Kozorog Marc-Antoine Sani Martina Lenarčič Živković Gregor Ilc Vesna Hodnik Frances Separovic Janez Plavec Gregor Anderluh 19F NMR studies provide insights into lipid membrane interactions of listeriolysin O, a pore forming toxin from Listeria monocytogenes
description	Abstract Listeria monocytogenes is a mammalian pathogen that causes gastroenteritis, miscarriages and infections of the central nervous system in immunocompromised individuals. Its main virulence factor is listeriolysin O (LLO), a pore-forming cholesterol-dependent cytolysin (CDC), which enables bacterial escape from the phagolysosome and contributes to bacterial pathogenicity. Details of cholesterol (Chol) recognition and membrane binding mechanisms by LLO are still not known. Here we used 19F-NMR spectroscopy in order to assess LLO-Chol interactions in solution and in a Chol-rich membrane environment. LLO has six tryptophan residues located in the region of the molecule that is first in contact with lipid membranes. 19F-LLO, which contained 5-fluoro-tryptophans, was prepared by using isotopic labelling in an E. coli expression system. Signals in the 19F-NMR spectrum of 19F-LLO were unambiguously assigned by using a series of single Trp → Phe point mutations. The results employing various cholesterol preparations in solution indicate that tryptophan residues are not directly involved in Chol binding in solution. However, significant chemical shift changes were observed upon LLO binding to Chol-rich membranes, highlighting the role of tryptophan residues in membrane interactions (W512) and oligomerisation (W189 and W489).
format	article
author	Mirijam Kozorog Marc-Antoine Sani Martina Lenarčič Živković Gregor Ilc Vesna Hodnik Frances Separovic Janez Plavec Gregor Anderluh
author_facet	Mirijam Kozorog Marc-Antoine Sani Martina Lenarčič Živković Gregor Ilc Vesna Hodnik Frances Separovic Janez Plavec Gregor Anderluh
author_sort	Mirijam Kozorog
title	19F NMR studies provide insights into lipid membrane interactions of listeriolysin O, a pore forming toxin from Listeria monocytogenes
title_short	19F NMR studies provide insights into lipid membrane interactions of listeriolysin O, a pore forming toxin from Listeria monocytogenes
title_full	19F NMR studies provide insights into lipid membrane interactions of listeriolysin O, a pore forming toxin from Listeria monocytogenes
title_fullStr	19F NMR studies provide insights into lipid membrane interactions of listeriolysin O, a pore forming toxin from Listeria monocytogenes
title_full_unstemmed	19F NMR studies provide insights into lipid membrane interactions of listeriolysin O, a pore forming toxin from Listeria monocytogenes
title_sort	19f nmr studies provide insights into lipid membrane interactions of listeriolysin o, a pore forming toxin from listeria monocytogenes
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/d6860f4192aa49c7bb6735ed309dc05f
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_version_	1718388091484372992

19F NMR studies provide insights into lipid membrane interactions of listeriolysin O, a pore forming toxin from Listeria monocytogenes

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