Identification of a membrane-bound prepore species clarifies the lytic mechanism of actinoporins

Autores
Morante, Kodo; Bellomio, Augusto; Gil Cartón. David; Redondo Morata, Lorena; Sot, Jesús; Scheuring, Simon; Valle, Mikel; González Mañas, Juan Manuel; Tsumoto, Kohuei; Caaveiro, José M. M.
Año de publicación
2016
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Pore-forming toxins (PFTs) are cytolytic proteins belonging to the molecular warfare apparatus of living organisms. The assembly of the functional transmembrane pore requires several intermediate steps ranging from a water-soluble monomeric species to the multimeric ensemble inserted in the cell membrane. The non-lytic oligomeric intermediate known as prepore plays an essential role in the mechanism of insertion of the class of β-PFTs. However, in the class of α-PFTs, like the actinoporins produced by sea anemones, evidence of membrane-bound prepores is still lacking. We have employed single-particle cryo-electron microscopy (cryo-EM) and atomic force microscopy to identify, for the first time, a prepore species of the actinoporin fragaceatoxin C bound to lipid vesicles. The size of the prepore coincides with that of the functional pore, except for the transmembrane region, which is absent in the prepore. Biochemical assays indicated that, in the prepore species, the N terminus is not inserted in the bilayer but is exposed to the aqueous solution. Our study reveals the structure of the prepore in actinoporins and highlights the role of structural intermediates for the formation of cytolytic pores by an α-PFT.
Fil: Morante, Kodo. Universidad del País Vasco; España. University of Tokyo; Japón
Fil: Bellomio, Augusto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto Superior de Investigaciones Biológicas. Universidad Nacional de Tucumán. Instituto Superior de Investigaciones Biológicas; Argentina. Universidad del País Vasco; España. Consejo Superior de Investigaciones Científicas; España
Fil: Gil Cartón. David. CICbiogune; España
Fil: Redondo Morata, Lorena. Inserm; Francia. Aix-Marseille Université; Francia
Fil: Sot, Jesús. Universidad del País Vasco; España. Consejo Superior de Investigaciones Científicas; España
Fil: Scheuring, Simon. Inserm; Francia. Aix-Marseille Université; Francia
Fil: Valle, Mikel. CICbiogune; España
Fil: González Mañas, Juan Manuel. Universidad del País Vasco; España. Consejo Superior de Investigaciones Científicas; España
Fil: Tsumoto, Kohuei. University of Tokyo; Japón
Fil: Caaveiro, José M. M.. University of Tokyo; Japón
Materia
Pore Forming Protein
Cytolysin
Lipid‐Protein Interaction
Protein Structure
Oligomerization
Atomic Force Microscopy
Lipid Vesicle
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/51217
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/51217
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Identification of a membrane-bound prepore species clarifies the lytic mechanism of actinoporinsMorante, KodoBellomio, AugustoGil Cartón. DavidRedondo Morata, LorenaSot, JesúsScheuring, SimonValle, MikelGonzález Mañas, Juan ManuelTsumoto, KohueiCaaveiro, José M. M.Pore Forming ProteinCytolysinLipid‐Protein InteractionProtein StructureOligomerizationAtomic Force MicroscopyLipid Vesiclehttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Pore-forming toxins (PFTs) are cytolytic proteins belonging to the molecular warfare apparatus of living organisms. The assembly of the functional transmembrane pore requires several intermediate steps ranging from a water-soluble monomeric species to the multimeric ensemble inserted in the cell membrane. The non-lytic oligomeric intermediate known as prepore plays an essential role in the mechanism of insertion of the class of β-PFTs. However, in the class of α-PFTs, like the actinoporins produced by sea anemones, evidence of membrane-bound prepores is still lacking. We have employed single-particle cryo-electron microscopy (cryo-EM) and atomic force microscopy to identify, for the first time, a prepore species of the actinoporin fragaceatoxin C bound to lipid vesicles. The size of the prepore coincides with that of the functional pore, except for the transmembrane region, which is absent in the prepore. Biochemical assays indicated that, in the prepore species, the N terminus is not inserted in the bilayer but is exposed to the aqueous solution. Our study reveals the structure of the prepore in actinoporins and highlights the role of structural intermediates for the formation of cytolytic pores by an α-PFT.Fil: Morante, Kodo. Universidad del País Vasco; España. University of Tokyo; JapónFil: Bellomio, Augusto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto Superior de Investigaciones Biológicas. Universidad Nacional de Tucumán. Instituto Superior de Investigaciones Biológicas; Argentina. Universidad del País Vasco; España. Consejo Superior de Investigaciones Científicas; EspañaFil: Gil Cartón. David. CICbiogune; EspañaFil: Redondo Morata, Lorena. Inserm; Francia. Aix-Marseille Université; FranciaFil: Sot, Jesús. Universidad del País Vasco; España. Consejo Superior de Investigaciones Científicas; EspañaFil: Scheuring, Simon. Inserm; Francia. Aix-Marseille Université; FranciaFil: Valle, Mikel. CICbiogune; EspañaFil: González Mañas, Juan Manuel. Universidad del País Vasco; España. Consejo Superior de Investigaciones Científicas; EspañaFil: Tsumoto, Kohuei. University of Tokyo; JapónFil: Caaveiro, José M. M.. University of Tokyo; JapónAmerican Society for Biochemistry and Molecular Biology2016-09info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/51217Morante, Kodo; Bellomio, Augusto; Gil Cartón. David; Redondo Morata, Lorena; Sot, Jesús; et al.; Identification of a membrane-bound prepore species clarifies the lytic mechanism of actinoporins; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 291; 37; 9-2016; 19210-192190021-9258CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.jbc.org/content/291/37/19210.fullinfo:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.M116.734053info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:18:40Zoai:ri.conicet.gov.ar:11336/51217instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:18:41.262CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Identification of a membrane-bound prepore species clarifies the lytic mechanism of actinoporins
title Identification of a membrane-bound prepore species clarifies the lytic mechanism of actinoporins
spellingShingle Identification of a membrane-bound prepore species clarifies the lytic mechanism of actinoporins
Morante, Kodo
Pore Forming Protein
Cytolysin
Lipid‐Protein Interaction
Protein Structure
Oligomerization
Atomic Force Microscopy
Lipid Vesicle
title_short Identification of a membrane-bound prepore species clarifies the lytic mechanism of actinoporins
title_full Identification of a membrane-bound prepore species clarifies the lytic mechanism of actinoporins
title_fullStr Identification of a membrane-bound prepore species clarifies the lytic mechanism of actinoporins
title_full_unstemmed Identification of a membrane-bound prepore species clarifies the lytic mechanism of actinoporins
title_sort Identification of a membrane-bound prepore species clarifies the lytic mechanism of actinoporins
dc.creator.none.fl_str_mv Morante, Kodo
Bellomio, Augusto
Gil Cartón. David
Redondo Morata, Lorena
Sot, Jesús
Scheuring, Simon
Valle, Mikel
González Mañas, Juan Manuel
Tsumoto, Kohuei
Caaveiro, José M. M.
author Morante, Kodo
author_facet Morante, Kodo
Bellomio, Augusto
Gil Cartón. David
Redondo Morata, Lorena
Sot, Jesús
Scheuring, Simon
Valle, Mikel
González Mañas, Juan Manuel
Tsumoto, Kohuei
Caaveiro, José M. M.
author_role author
author2 Bellomio, Augusto
Gil Cartón. David
Redondo Morata, Lorena
Sot, Jesús
Scheuring, Simon
Valle, Mikel
González Mañas, Juan Manuel
Tsumoto, Kohuei
Caaveiro, José M. M.
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Pore Forming Protein
Cytolysin
Lipid‐Protein Interaction
Protein Structure
Oligomerization
Atomic Force Microscopy
Lipid Vesicle
topic Pore Forming Protein
Cytolysin
Lipid‐Protein Interaction
Protein Structure
Oligomerization
Atomic Force Microscopy
Lipid Vesicle
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Pore-forming toxins (PFTs) are cytolytic proteins belonging to the molecular warfare apparatus of living organisms. The assembly of the functional transmembrane pore requires several intermediate steps ranging from a water-soluble monomeric species to the multimeric ensemble inserted in the cell membrane. The non-lytic oligomeric intermediate known as prepore plays an essential role in the mechanism of insertion of the class of β-PFTs. However, in the class of α-PFTs, like the actinoporins produced by sea anemones, evidence of membrane-bound prepores is still lacking. We have employed single-particle cryo-electron microscopy (cryo-EM) and atomic force microscopy to identify, for the first time, a prepore species of the actinoporin fragaceatoxin C bound to lipid vesicles. The size of the prepore coincides with that of the functional pore, except for the transmembrane region, which is absent in the prepore. Biochemical assays indicated that, in the prepore species, the N terminus is not inserted in the bilayer but is exposed to the aqueous solution. Our study reveals the structure of the prepore in actinoporins and highlights the role of structural intermediates for the formation of cytolytic pores by an α-PFT.
Fil: Morante, Kodo. Universidad del País Vasco; España. University of Tokyo; Japón
Fil: Bellomio, Augusto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto Superior de Investigaciones Biológicas. Universidad Nacional de Tucumán. Instituto Superior de Investigaciones Biológicas; Argentina. Universidad del País Vasco; España. Consejo Superior de Investigaciones Científicas; España
Fil: Gil Cartón. David. CICbiogune; España
Fil: Redondo Morata, Lorena. Inserm; Francia. Aix-Marseille Université; Francia
Fil: Sot, Jesús. Universidad del País Vasco; España. Consejo Superior de Investigaciones Científicas; España
Fil: Scheuring, Simon. Inserm; Francia. Aix-Marseille Université; Francia
Fil: Valle, Mikel. CICbiogune; España
Fil: González Mañas, Juan Manuel. Universidad del País Vasco; España. Consejo Superior de Investigaciones Científicas; España
Fil: Tsumoto, Kohuei. University of Tokyo; Japón
Fil: Caaveiro, José M. M.. University of Tokyo; Japón
description Pore-forming toxins (PFTs) are cytolytic proteins belonging to the molecular warfare apparatus of living organisms. The assembly of the functional transmembrane pore requires several intermediate steps ranging from a water-soluble monomeric species to the multimeric ensemble inserted in the cell membrane. The non-lytic oligomeric intermediate known as prepore plays an essential role in the mechanism of insertion of the class of β-PFTs. However, in the class of α-PFTs, like the actinoporins produced by sea anemones, evidence of membrane-bound prepores is still lacking. We have employed single-particle cryo-electron microscopy (cryo-EM) and atomic force microscopy to identify, for the first time, a prepore species of the actinoporin fragaceatoxin C bound to lipid vesicles. The size of the prepore coincides with that of the functional pore, except for the transmembrane region, which is absent in the prepore. Biochemical assays indicated that, in the prepore species, the N terminus is not inserted in the bilayer but is exposed to the aqueous solution. Our study reveals the structure of the prepore in actinoporins and highlights the role of structural intermediates for the formation of cytolytic pores by an α-PFT.
publishDate 2016
dc.date.none.fl_str_mv 2016-09
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/51217
Morante, Kodo; Bellomio, Augusto; Gil Cartón. David; Redondo Morata, Lorena; Sot, Jesús; et al.; Identification of a membrane-bound prepore species clarifies the lytic mechanism of actinoporins; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 291; 37; 9-2016; 19210-19219
0021-9258
CONICET Digital
CONICET
url http://hdl.handle.net/11336/51217
identifier_str_mv Morante, Kodo; Bellomio, Augusto; Gil Cartón. David; Redondo Morata, Lorena; Sot, Jesús; et al.; Identification of a membrane-bound prepore species clarifies the lytic mechanism of actinoporins; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 291; 37; 9-2016; 19210-19219
0021-9258
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.jbc.org/content/291/37/19210.full
info:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.M116.734053
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.070432

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