Coupling between GABAA-R ligand-binding activity and membrane organization in β-cyclodextrin-treated synaptosomal membranes from bovine brain cortex: new insights from EPR experime...

Autores
Turina, Anahi del Valle; Schreier, Shirley; Perillo, Maria Angelica
Año de publicación
2012
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Correlations between GABAA-receptor (GABAA-R) activity and molecular organization of synaptosomal membranes (SM) were studied along the protocol for cholesterol (Cho) extraction with b-cyclodextrin (b-CD). The mere pre-incubation (PI) at 37ºC accompanying the b-CD treatment was an underlying source of perturbations increasing [3H]-FNZ maximal binding (70%) and Kd (38%), plus a stiffening of SMs’ hydrocarbon core region. The latter was inferred from an increased compressibility modulus (K) of SM-derived Langmuir films, a blue-shifted DPH fluorescence emission spectrum and the hysteresis in DPH fluorescence anisotropy (ADPH) in SMs submitted to a heating-cooling cycle (4ºC-37ºC-4ºC) with ADPH,heatingFil: Turina, Anahi del Valle. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina
Fil: Schreier, Shirley. Universidade de Sao Paulo; Brasil
Fil: Perillo, Maria Angelica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina
Materia
Cyclodextrin
Gabaa Receptor
Epr
Fluorescence Anisotropy
Langmuir Films
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:11336/42608
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/42608
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Coupling between GABAA-R ligand-binding activity and membrane organization in β-cyclodextrin-treated synaptosomal membranes from bovine brain cortex: new insights from EPR experimentsTurina, Anahi del ValleSchreier, ShirleyPerillo, Maria AngelicaCyclodextrinGabaa ReceptorEprFluorescence AnisotropyLangmuir Filmshttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Correlations between GABAA-receptor (GABAA-R) activity and molecular organization of synaptosomal membranes (SM) were studied along the protocol for cholesterol (Cho) extraction with b-cyclodextrin (b-CD). The mere pre-incubation (PI) at 37ºC accompanying the b-CD treatment was an underlying source of perturbations increasing [3H]-FNZ maximal binding (70%) and Kd (38%), plus a stiffening of SMs’ hydrocarbon core region. The latter was inferred from an increased compressibility modulus (K) of SM-derived Langmuir films, a blue-shifted DPH fluorescence emission spectrum and the hysteresis in DPH fluorescence anisotropy (ADPH) in SMs submitted to a heating-cooling cycle (4ºC-37ºC-4ºC) with ADPH,heating<ADPH,cooling. Compared with PI-samples, the b-CD treatment reduced Bmax by 5% which correlated with a 45%-decrement in the relative Cho content of SM, a decrease in K and in the order parameter in the EPR spectrum of a lipid spin probe labeled at C5 (5-SASL), and significantly increased ATMA-DPH. PI, but not b-CD treatment, could affect the binding affinity. EPR spectra of 5-SASL complexes with b-CD-, SM-partitioned and free in solution showed that, contrary to what is usually assumed, b-CD is not completely eliminated from the system through centrifugation washings. It was concluded that, b-CD treatment involves effects of at least three different types of events affecting membrane organization: a) effect of PI on membrane annealing, b) effect of residual b-CD on SM organization and c) Cho depletion. Consequently, molecular stiffness increases within the membrane core and decreases near the polar head groups, leading to a net increase in GABAA-R density, relative to untreated samples.Fil: Turina, Anahi del Valle. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; ArgentinaFil: Schreier, Shirley. Universidade de Sao Paulo; BrasilFil: Perillo, Maria Angelica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; ArgentinaHumana Press2012-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/42608Turina, Anahi del Valle; Schreier, Shirley; Perillo, Maria Angelica; Coupling between GABAA-R ligand-binding activity and membrane organization in β-cyclodextrin-treated synaptosomal membranes from bovine brain cortex: new insights from EPR experiments; Humana Press; Cell Biochemistry and Biophysics; 63; 1; 2-2012; 17-331085-91951559-0283CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://link.springer.com/article/10.1007%2Fs12013-012-9338-1info:eu-repo/semantics/altIdentifier/doi/10.1007/s12013-012-9338-1info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:27:43Zoai:ri.conicet.gov.ar:11336/42608instacron: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 10:27:43.752CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Coupling between GABAA-R ligand-binding activity and membrane organization in β-cyclodextrin-treated synaptosomal membranes from bovine brain cortex: new insights from EPR experiments
title Coupling between GABAA-R ligand-binding activity and membrane organization in β-cyclodextrin-treated synaptosomal membranes from bovine brain cortex: new insights from EPR experiments
spellingShingle Coupling between GABAA-R ligand-binding activity and membrane organization in β-cyclodextrin-treated synaptosomal membranes from bovine brain cortex: new insights from EPR experiments
Turina, Anahi del Valle
Cyclodextrin
Gabaa Receptor
Epr
Fluorescence Anisotropy
Langmuir Films
title_short Coupling between GABAA-R ligand-binding activity and membrane organization in β-cyclodextrin-treated synaptosomal membranes from bovine brain cortex: new insights from EPR experiments
title_full Coupling between GABAA-R ligand-binding activity and membrane organization in β-cyclodextrin-treated synaptosomal membranes from bovine brain cortex: new insights from EPR experiments
title_fullStr Coupling between GABAA-R ligand-binding activity and membrane organization in β-cyclodextrin-treated synaptosomal membranes from bovine brain cortex: new insights from EPR experiments
title_full_unstemmed Coupling between GABAA-R ligand-binding activity and membrane organization in β-cyclodextrin-treated synaptosomal membranes from bovine brain cortex: new insights from EPR experiments
title_sort Coupling between GABAA-R ligand-binding activity and membrane organization in β-cyclodextrin-treated synaptosomal membranes from bovine brain cortex: new insights from EPR experiments
dc.creator.none.fl_str_mv Turina, Anahi del Valle
Schreier, Shirley
Perillo, Maria Angelica
author Turina, Anahi del Valle
author_facet Turina, Anahi del Valle
Schreier, Shirley
Perillo, Maria Angelica
author_role author
author2 Schreier, Shirley
Perillo, Maria Angelica
author2_role author
author
dc.subject.none.fl_str_mv Cyclodextrin
Gabaa Receptor
Epr
Fluorescence Anisotropy
Langmuir Films
topic Cyclodextrin
Gabaa Receptor
Epr
Fluorescence Anisotropy
Langmuir Films
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Correlations between GABAA-receptor (GABAA-R) activity and molecular organization of synaptosomal membranes (SM) were studied along the protocol for cholesterol (Cho) extraction with b-cyclodextrin (b-CD). The mere pre-incubation (PI) at 37ºC accompanying the b-CD treatment was an underlying source of perturbations increasing [3H]-FNZ maximal binding (70%) and Kd (38%), plus a stiffening of SMs’ hydrocarbon core region. The latter was inferred from an increased compressibility modulus (K) of SM-derived Langmuir films, a blue-shifted DPH fluorescence emission spectrum and the hysteresis in DPH fluorescence anisotropy (ADPH) in SMs submitted to a heating-cooling cycle (4ºC-37ºC-4ºC) with ADPH,heating<ADPH,cooling. Compared with PI-samples, the b-CD treatment reduced Bmax by 5% which correlated with a 45%-decrement in the relative Cho content of SM, a decrease in K and in the order parameter in the EPR spectrum of a lipid spin probe labeled at C5 (5-SASL), and significantly increased ATMA-DPH. PI, but not b-CD treatment, could affect the binding affinity. EPR spectra of 5-SASL complexes with b-CD-, SM-partitioned and free in solution showed that, contrary to what is usually assumed, b-CD is not completely eliminated from the system through centrifugation washings. It was concluded that, b-CD treatment involves effects of at least three different types of events affecting membrane organization: a) effect of PI on membrane annealing, b) effect of residual b-CD on SM organization and c) Cho depletion. Consequently, molecular stiffness increases within the membrane core and decreases near the polar head groups, leading to a net increase in GABAA-R density, relative to untreated samples.
Fil: Turina, Anahi del Valle. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina
Fil: Schreier, Shirley. Universidade de Sao Paulo; Brasil
Fil: Perillo, Maria Angelica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina
description Correlations between GABAA-receptor (GABAA-R) activity and molecular organization of synaptosomal membranes (SM) were studied along the protocol for cholesterol (Cho) extraction with b-cyclodextrin (b-CD). The mere pre-incubation (PI) at 37ºC accompanying the b-CD treatment was an underlying source of perturbations increasing [3H]-FNZ maximal binding (70%) and Kd (38%), plus a stiffening of SMs’ hydrocarbon core region. The latter was inferred from an increased compressibility modulus (K) of SM-derived Langmuir films, a blue-shifted DPH fluorescence emission spectrum and the hysteresis in DPH fluorescence anisotropy (ADPH) in SMs submitted to a heating-cooling cycle (4ºC-37ºC-4ºC) with ADPH,heating<ADPH,cooling. Compared with PI-samples, the b-CD treatment reduced Bmax by 5% which correlated with a 45%-decrement in the relative Cho content of SM, a decrease in K and in the order parameter in the EPR spectrum of a lipid spin probe labeled at C5 (5-SASL), and significantly increased ATMA-DPH. PI, but not b-CD treatment, could affect the binding affinity. EPR spectra of 5-SASL complexes with b-CD-, SM-partitioned and free in solution showed that, contrary to what is usually assumed, b-CD is not completely eliminated from the system through centrifugation washings. It was concluded that, b-CD treatment involves effects of at least three different types of events affecting membrane organization: a) effect of PI on membrane annealing, b) effect of residual b-CD on SM organization and c) Cho depletion. Consequently, molecular stiffness increases within the membrane core and decreases near the polar head groups, leading to a net increase in GABAA-R density, relative to untreated samples.
publishDate 2012
dc.date.none.fl_str_mv 2012-02
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/11336/42608
Turina, Anahi del Valle; Schreier, Shirley; Perillo, Maria Angelica; Coupling between GABAA-R ligand-binding activity and membrane organization in β-cyclodextrin-treated synaptosomal membranes from bovine brain cortex: new insights from EPR experiments; Humana Press; Cell Biochemistry and Biophysics; 63; 1; 2-2012; 17-33
1085-9195
1559-0283
CONICET Digital
CONICET
url http://hdl.handle.net/11336/42608
identifier_str_mv Turina, Anahi del Valle; Schreier, Shirley; Perillo, Maria Angelica; Coupling between GABAA-R ligand-binding activity and membrane organization in β-cyclodextrin-treated synaptosomal membranes from bovine brain cortex: new insights from EPR experiments; Humana Press; Cell Biochemistry and Biophysics; 63; 1; 2-2012; 17-33
1085-9195
1559-0283
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://link.springer.com/article/10.1007%2Fs12013-012-9338-1
info:eu-repo/semantics/altIdentifier/doi/10.1007/s12013-012-9338-1
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 Humana Press
publisher.none.fl_str_mv Humana Press
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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