Lipid bilayer-atomic force microscopy combined platform records simultaneous electrical and topological changes of the TRP channel Polycystin-2 (TRPP2)

Autores
Lal S; Scarinci, María Noelia; Pérez, Paula Luciana; Cantiello, Horacio Fabio; Cantero, Maria del Rocio
Año de publicación
2018
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Ion channels are transmembrane proteins that mediate ion transport across biological membranes. Ion channel function is traditionally characterized by electrical parameters acquired with techniques such as patch-clamping and reconstitution in lipid bilayer membranes (BLM) that provide relevant information such as ionic conductance, selectivity, and gating properties. High resolution structural information of ion channels however, requires independent technologies, of which atomic force microscopy (AFM) is the only one that provides topological features of single functional channel proteins in their native environments. To date practically no data exist on direct correlations between electrical features and topological parameters from functional single channel complexes. Here, we report the design and construction of a BLM reconstitution microchamber that supports the simultaneous recording of electrical currents and AFM imaging from single channel complexes. As proof-of-principle, we tested the technique on polycystin-2 (PC2, TRPP2), a TRP channel family member from which we had previously elucidated its tetrameric topology by AFM imaging, and single channel currents by the BLM technique. The experimental setup provided direct structural-functional correlates from PC2 single channel complexes that disclosed novel topological changes between the closed and open sub-conductance states of the functional channel, namely, an inverse correlation between conductance and height of the channel. Unexpectedly, we also disclosed intrinsic PC2 mechanosensitivity in response to external forces. The platform provides a suitable means of accessing topological information to correlate with ion channel electrical parameters essential to understand the physiology of these transmembrane proteins.
Fil: Lal S. Massachusetts General Hospital And Harvard Medical School;
Fil: Scarinci, María Noelia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Pérez, Paula Luciana. Universidad de Buenos Aires; Argentina
Fil: Cantiello, Horacio Fabio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Cantero, Maria del Rocio. Universidad de Buenos Aires. Facultad de Odontología. Cátedra de Biofísica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Materia
POLYCYSTIN-2
AFM
ELECTROPHYSIOLOGY
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:11336/81390

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network_name_str CONICET Digital (CONICET)
spelling Lipid bilayer-atomic force microscopy combined platform records simultaneous electrical and topological changes of the TRP channel Polycystin-2 (TRPP2)Lal SScarinci, María NoeliaPérez, Paula LucianaCantiello, Horacio FabioCantero, Maria del RocioPOLYCYSTIN-2AFMELECTROPHYSIOLOGYhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Ion channels are transmembrane proteins that mediate ion transport across biological membranes. Ion channel function is traditionally characterized by electrical parameters acquired with techniques such as patch-clamping and reconstitution in lipid bilayer membranes (BLM) that provide relevant information such as ionic conductance, selectivity, and gating properties. High resolution structural information of ion channels however, requires independent technologies, of which atomic force microscopy (AFM) is the only one that provides topological features of single functional channel proteins in their native environments. To date practically no data exist on direct correlations between electrical features and topological parameters from functional single channel complexes. Here, we report the design and construction of a BLM reconstitution microchamber that supports the simultaneous recording of electrical currents and AFM imaging from single channel complexes. As proof-of-principle, we tested the technique on polycystin-2 (PC2, TRPP2), a TRP channel family member from which we had previously elucidated its tetrameric topology by AFM imaging, and single channel currents by the BLM technique. The experimental setup provided direct structural-functional correlates from PC2 single channel complexes that disclosed novel topological changes between the closed and open sub-conductance states of the functional channel, namely, an inverse correlation between conductance and height of the channel. Unexpectedly, we also disclosed intrinsic PC2 mechanosensitivity in response to external forces. The platform provides a suitable means of accessing topological information to correlate with ion channel electrical parameters essential to understand the physiology of these transmembrane proteins.Fil: Lal S. Massachusetts General Hospital And Harvard Medical School;Fil: Scarinci, María Noelia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; ArgentinaFil: Pérez, Paula Luciana. Universidad de Buenos Aires; ArgentinaFil: Cantiello, Horacio Fabio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; ArgentinaFil: Cantero, Maria del Rocio. Universidad de Buenos Aires. Facultad de Odontología. Cátedra de Biofísica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; ArgentinaPublic Library of Science2018-08info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/81390Lal S; Scarinci, María Noelia; Pérez, Paula Luciana; Cantiello, Horacio Fabio; Cantero, Maria del Rocio; Lipid bilayer-atomic force microscopy combined platform records simultaneous electrical and topological changes of the TRP channel Polycystin-2 (TRPP2); Public Library of Science; Plos One; 8-2018; 1-171932-6203CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1371/journal.pone.0202029info:eu-repo/semantics/altIdentifier/url/https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0202029info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T12:00:37Zoai:ri.conicet.gov.ar:11336/81390instacron: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:34982025-10-22 12:00:38.094CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Lipid bilayer-atomic force microscopy combined platform records simultaneous electrical and topological changes of the TRP channel Polycystin-2 (TRPP2)
title Lipid bilayer-atomic force microscopy combined platform records simultaneous electrical and topological changes of the TRP channel Polycystin-2 (TRPP2)
spellingShingle Lipid bilayer-atomic force microscopy combined platform records simultaneous electrical and topological changes of the TRP channel Polycystin-2 (TRPP2)
Lal S
POLYCYSTIN-2
AFM
ELECTROPHYSIOLOGY
title_short Lipid bilayer-atomic force microscopy combined platform records simultaneous electrical and topological changes of the TRP channel Polycystin-2 (TRPP2)
title_full Lipid bilayer-atomic force microscopy combined platform records simultaneous electrical and topological changes of the TRP channel Polycystin-2 (TRPP2)
title_fullStr Lipid bilayer-atomic force microscopy combined platform records simultaneous electrical and topological changes of the TRP channel Polycystin-2 (TRPP2)
title_full_unstemmed Lipid bilayer-atomic force microscopy combined platform records simultaneous electrical and topological changes of the TRP channel Polycystin-2 (TRPP2)
title_sort Lipid bilayer-atomic force microscopy combined platform records simultaneous electrical and topological changes of the TRP channel Polycystin-2 (TRPP2)
dc.creator.none.fl_str_mv Lal S
Scarinci, María Noelia
Pérez, Paula Luciana
Cantiello, Horacio Fabio
Cantero, Maria del Rocio
author Lal S
author_facet Lal S
Scarinci, María Noelia
Pérez, Paula Luciana
Cantiello, Horacio Fabio
Cantero, Maria del Rocio
author_role author
author2 Scarinci, María Noelia
Pérez, Paula Luciana
Cantiello, Horacio Fabio
Cantero, Maria del Rocio
author2_role author
author
author
author
dc.subject.none.fl_str_mv POLYCYSTIN-2
AFM
ELECTROPHYSIOLOGY
topic POLYCYSTIN-2
AFM
ELECTROPHYSIOLOGY
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Ion channels are transmembrane proteins that mediate ion transport across biological membranes. Ion channel function is traditionally characterized by electrical parameters acquired with techniques such as patch-clamping and reconstitution in lipid bilayer membranes (BLM) that provide relevant information such as ionic conductance, selectivity, and gating properties. High resolution structural information of ion channels however, requires independent technologies, of which atomic force microscopy (AFM) is the only one that provides topological features of single functional channel proteins in their native environments. To date practically no data exist on direct correlations between electrical features and topological parameters from functional single channel complexes. Here, we report the design and construction of a BLM reconstitution microchamber that supports the simultaneous recording of electrical currents and AFM imaging from single channel complexes. As proof-of-principle, we tested the technique on polycystin-2 (PC2, TRPP2), a TRP channel family member from which we had previously elucidated its tetrameric topology by AFM imaging, and single channel currents by the BLM technique. The experimental setup provided direct structural-functional correlates from PC2 single channel complexes that disclosed novel topological changes between the closed and open sub-conductance states of the functional channel, namely, an inverse correlation between conductance and height of the channel. Unexpectedly, we also disclosed intrinsic PC2 mechanosensitivity in response to external forces. The platform provides a suitable means of accessing topological information to correlate with ion channel electrical parameters essential to understand the physiology of these transmembrane proteins.
Fil: Lal S. Massachusetts General Hospital And Harvard Medical School;
Fil: Scarinci, María Noelia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Pérez, Paula Luciana. Universidad de Buenos Aires; Argentina
Fil: Cantiello, Horacio Fabio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Cantero, Maria del Rocio. Universidad de Buenos Aires. Facultad de Odontología. Cátedra de Biofísica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
description Ion channels are transmembrane proteins that mediate ion transport across biological membranes. Ion channel function is traditionally characterized by electrical parameters acquired with techniques such as patch-clamping and reconstitution in lipid bilayer membranes (BLM) that provide relevant information such as ionic conductance, selectivity, and gating properties. High resolution structural information of ion channels however, requires independent technologies, of which atomic force microscopy (AFM) is the only one that provides topological features of single functional channel proteins in their native environments. To date practically no data exist on direct correlations between electrical features and topological parameters from functional single channel complexes. Here, we report the design and construction of a BLM reconstitution microchamber that supports the simultaneous recording of electrical currents and AFM imaging from single channel complexes. As proof-of-principle, we tested the technique on polycystin-2 (PC2, TRPP2), a TRP channel family member from which we had previously elucidated its tetrameric topology by AFM imaging, and single channel currents by the BLM technique. The experimental setup provided direct structural-functional correlates from PC2 single channel complexes that disclosed novel topological changes between the closed and open sub-conductance states of the functional channel, namely, an inverse correlation between conductance and height of the channel. Unexpectedly, we also disclosed intrinsic PC2 mechanosensitivity in response to external forces. The platform provides a suitable means of accessing topological information to correlate with ion channel electrical parameters essential to understand the physiology of these transmembrane proteins.
publishDate 2018
dc.date.none.fl_str_mv 2018-08
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/11336/81390
Lal S; Scarinci, María Noelia; Pérez, Paula Luciana; Cantiello, Horacio Fabio; Cantero, Maria del Rocio; Lipid bilayer-atomic force microscopy combined platform records simultaneous electrical and topological changes of the TRP channel Polycystin-2 (TRPP2); Public Library of Science; Plos One; 8-2018; 1-17
1932-6203
CONICET Digital
CONICET
url http://hdl.handle.net/11336/81390
identifier_str_mv Lal S; Scarinci, María Noelia; Pérez, Paula Luciana; Cantiello, Horacio Fabio; Cantero, Maria del Rocio; Lipid bilayer-atomic force microscopy combined platform records simultaneous electrical and topological changes of the TRP channel Polycystin-2 (TRPP2); Public Library of Science; Plos One; 8-2018; 1-17
1932-6203
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1371/journal.pone.0202029
info:eu-repo/semantics/altIdentifier/url/https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0202029
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
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Public Library of Science
publisher.none.fl_str_mv Public Library of Science
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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