Mfn2-dependent fusion pathway of PE-enriched micron-sized vesicles

Autores
Peñalva, Daniel Alejandro; Monnappa, Ajay K.; Natale, Paolo; López Montero, Iván
Año de publicación
2024
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Mitofusins (Mfn1 and Mfn2) are the mitochondrial outer-membrane fusion proteins in mammals and belong to the dynamin superfamily of multidomain GTPases. Recent structural studies of truncated variants lacking alpha helical transmembrane domains suggested that Mfns dimerize to promote the approximation and the fusion of the mitochondrial outer membranes upon the hydrolysis of guanine 5′-triphosphate disodium salt (GTP). However, next to the presence of GTP, the fusion activity seems to require multiple regulatory factors that control the dynamics and kinetics of mitochondrial fusion through the formation of Mfn1–Mfn2 heterodimers. Here, we purified and reconstituted the full-length murine Mfn2 protein into giant unilamellar vesicles (GUVs) with different lipid compositions. The incubation with GTP resulted in the fusion of Mfn2-GUVs. High-speed video-microscopy showed that the Mfn2-dependent membrane fusion pathway progressed through a zipper mechanism where the formation and growth of an adhesion patch eventually led to the formation of a membrane opening at the rim of the septum. The presence of physiological concentration (up to 30 mol%) of dioleoyl-phosphatidylethanolamine (DOPE) was shown to be a requisite to observe GTP-induced Mfn2-dependent fusion. Our observations show that Mfn2 alone can promote the fusion of micron-sized DOPE-enriched Vesicles without the requirement of regulatory cofactors, such as membrane curvature, or the assistance of other proteins.
Fil: Peñalva, Daniel Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Monnappa, Ajay K.. Instituto de Investigación Biomédica Hospital Doce de Octubre; España
Fil: Natale, Paolo. Universidad Complutense de Madrid; España
Fil: López Montero, Iván. Universidad Complutense de Madrid; España
Materia
MEMBRANE FUSION
GIANT UNILAMELLAR VESICLES
MITOCHONDRIAL DYNAMICS
MITOFUSIN 2
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/244690
Acceder
id CONICETDig_05fb1cd3b20f0a89493b5e8224965216
oai_identifier_str oai:ri.conicet.gov.ar:11336/244690
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Mfn2-dependent fusion pathway of PE-enriched micron-sized vesiclesPeñalva, Daniel AlejandroMonnappa, Ajay K.Natale, PaoloLópez Montero, IvánMEMBRANE FUSIONGIANT UNILAMELLAR VESICLESMITOCHONDRIAL DYNAMICSMITOFUSIN 2https://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Mitofusins (Mfn1 and Mfn2) are the mitochondrial outer-membrane fusion proteins in mammals and belong to the dynamin superfamily of multidomain GTPases. Recent structural studies of truncated variants lacking alpha helical transmembrane domains suggested that Mfns dimerize to promote the approximation and the fusion of the mitochondrial outer membranes upon the hydrolysis of guanine 5′-triphosphate disodium salt (GTP). However, next to the presence of GTP, the fusion activity seems to require multiple regulatory factors that control the dynamics and kinetics of mitochondrial fusion through the formation of Mfn1–Mfn2 heterodimers. Here, we purified and reconstituted the full-length murine Mfn2 protein into giant unilamellar vesicles (GUVs) with different lipid compositions. The incubation with GTP resulted in the fusion of Mfn2-GUVs. High-speed video-microscopy showed that the Mfn2-dependent membrane fusion pathway progressed through a zipper mechanism where the formation and growth of an adhesion patch eventually led to the formation of a membrane opening at the rim of the septum. The presence of physiological concentration (up to 30 mol%) of dioleoyl-phosphatidylethanolamine (DOPE) was shown to be a requisite to observe GTP-induced Mfn2-dependent fusion. Our observations show that Mfn2 alone can promote the fusion of micron-sized DOPE-enriched Vesicles without the requirement of regulatory cofactors, such as membrane curvature, or the assistance of other proteins.Fil: Peñalva, Daniel Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaFil: Monnappa, Ajay K.. Instituto de Investigación Biomédica Hospital Doce de Octubre; EspañaFil: Natale, Paolo. Universidad Complutense de Madrid; EspañaFil: López Montero, Iván. Universidad Complutense de Madrid; EspañaNational Academy of Sciences2024-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/244690Peñalva, Daniel Alejandro; Monnappa, Ajay K.; Natale, Paolo; López Montero, Iván; Mfn2-dependent fusion pathway of PE-enriched micron-sized vesicles; National Academy of Sciences; Proceedings of the National Academy of Sciences of The United States of America; 121; 30; 7-2024; 1-120027-8424CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://pnas.org/doi/10.1073/pnas.2313609121info:eu-repo/semantics/altIdentifier/doi/10.1073/pnas.2313609121info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:29:28Zoai:ri.conicet.gov.ar:11336/244690instacron: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 10:29:29.141CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Mfn2-dependent fusion pathway of PE-enriched micron-sized vesicles
title Mfn2-dependent fusion pathway of PE-enriched micron-sized vesicles
spellingShingle Mfn2-dependent fusion pathway of PE-enriched micron-sized vesicles
Peñalva, Daniel Alejandro
MEMBRANE FUSION
GIANT UNILAMELLAR VESICLES
MITOCHONDRIAL DYNAMICS
MITOFUSIN 2
title_short Mfn2-dependent fusion pathway of PE-enriched micron-sized vesicles
title_full Mfn2-dependent fusion pathway of PE-enriched micron-sized vesicles
title_fullStr Mfn2-dependent fusion pathway of PE-enriched micron-sized vesicles
title_full_unstemmed Mfn2-dependent fusion pathway of PE-enriched micron-sized vesicles
title_sort Mfn2-dependent fusion pathway of PE-enriched micron-sized vesicles
dc.creator.none.fl_str_mv Peñalva, Daniel Alejandro
Monnappa, Ajay K.
Natale, Paolo
López Montero, Iván
author Peñalva, Daniel Alejandro
author_facet Peñalva, Daniel Alejandro
Monnappa, Ajay K.
Natale, Paolo
López Montero, Iván
author_role author
author2 Monnappa, Ajay K.
Natale, Paolo
López Montero, Iván
author2_role author
author
author
dc.subject.none.fl_str_mv MEMBRANE FUSION
GIANT UNILAMELLAR VESICLES
MITOCHONDRIAL DYNAMICS
MITOFUSIN 2
topic MEMBRANE FUSION
GIANT UNILAMELLAR VESICLES
MITOCHONDRIAL DYNAMICS
MITOFUSIN 2
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Mitofusins (Mfn1 and Mfn2) are the mitochondrial outer-membrane fusion proteins in mammals and belong to the dynamin superfamily of multidomain GTPases. Recent structural studies of truncated variants lacking alpha helical transmembrane domains suggested that Mfns dimerize to promote the approximation and the fusion of the mitochondrial outer membranes upon the hydrolysis of guanine 5′-triphosphate disodium salt (GTP). However, next to the presence of GTP, the fusion activity seems to require multiple regulatory factors that control the dynamics and kinetics of mitochondrial fusion through the formation of Mfn1–Mfn2 heterodimers. Here, we purified and reconstituted the full-length murine Mfn2 protein into giant unilamellar vesicles (GUVs) with different lipid compositions. The incubation with GTP resulted in the fusion of Mfn2-GUVs. High-speed video-microscopy showed that the Mfn2-dependent membrane fusion pathway progressed through a zipper mechanism where the formation and growth of an adhesion patch eventually led to the formation of a membrane opening at the rim of the septum. The presence of physiological concentration (up to 30 mol%) of dioleoyl-phosphatidylethanolamine (DOPE) was shown to be a requisite to observe GTP-induced Mfn2-dependent fusion. Our observations show that Mfn2 alone can promote the fusion of micron-sized DOPE-enriched Vesicles without the requirement of regulatory cofactors, such as membrane curvature, or the assistance of other proteins.
Fil: Peñalva, Daniel Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Monnappa, Ajay K.. Instituto de Investigación Biomédica Hospital Doce de Octubre; España
Fil: Natale, Paolo. Universidad Complutense de Madrid; España
Fil: López Montero, Iván. Universidad Complutense de Madrid; España
description Mitofusins (Mfn1 and Mfn2) are the mitochondrial outer-membrane fusion proteins in mammals and belong to the dynamin superfamily of multidomain GTPases. Recent structural studies of truncated variants lacking alpha helical transmembrane domains suggested that Mfns dimerize to promote the approximation and the fusion of the mitochondrial outer membranes upon the hydrolysis of guanine 5′-triphosphate disodium salt (GTP). However, next to the presence of GTP, the fusion activity seems to require multiple regulatory factors that control the dynamics and kinetics of mitochondrial fusion through the formation of Mfn1–Mfn2 heterodimers. Here, we purified and reconstituted the full-length murine Mfn2 protein into giant unilamellar vesicles (GUVs) with different lipid compositions. The incubation with GTP resulted in the fusion of Mfn2-GUVs. High-speed video-microscopy showed that the Mfn2-dependent membrane fusion pathway progressed through a zipper mechanism where the formation and growth of an adhesion patch eventually led to the formation of a membrane opening at the rim of the septum. The presence of physiological concentration (up to 30 mol%) of dioleoyl-phosphatidylethanolamine (DOPE) was shown to be a requisite to observe GTP-induced Mfn2-dependent fusion. Our observations show that Mfn2 alone can promote the fusion of micron-sized DOPE-enriched Vesicles without the requirement of regulatory cofactors, such as membrane curvature, or the assistance of other proteins.
publishDate 2024
dc.date.none.fl_str_mv 2024-07
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/11336/244690
Peñalva, Daniel Alejandro; Monnappa, Ajay K.; Natale, Paolo; López Montero, Iván; Mfn2-dependent fusion pathway of PE-enriched micron-sized vesicles; National Academy of Sciences; Proceedings of the National Academy of Sciences of The United States of America; 121; 30; 7-2024; 1-12
0027-8424
CONICET Digital
CONICET
url http://hdl.handle.net/11336/244690
identifier_str_mv Peñalva, Daniel Alejandro; Monnappa, Ajay K.; Natale, Paolo; López Montero, Iván; Mfn2-dependent fusion pathway of PE-enriched micron-sized vesicles; National Academy of Sciences; Proceedings of the National Academy of Sciences of The United States of America; 121; 30; 7-2024; 1-12
0027-8424
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://pnas.org/doi/10.1073/pnas.2313609121
info:eu-repo/semantics/altIdentifier/doi/10.1073/pnas.2313609121
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv National Academy of Sciences
publisher.none.fl_str_mv National Academy of Sciences
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
_version_ 1844614301489299456
score 13.070432

Publicaciones similares