Resolution of complex fluorescence spectra of lipids and nicotinic acetylcholine receptor by multivariate analysis reveals protein-mediated effects on the receptor's immediate lipi...

Autores
Wenz, Jorge Javier; Barrantes, Francisco Jose
Año de publicación
2008
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Analysis of fluorescent spectra from comp lex biological systems containing various fluorescent probes with overlapping emissi on bands is a challenging task. Valuable information can be extracted from the full spectra, however, by using multivariate analysis (MA) of measurements at differen t wavelengths. We a pplied MA to spectral data of purified Torpedo nicotinic acetylcholine receptor (AChR) protein reconstituted into liposomes made up of diol eoylphosphatidic acid (DOPA) and dioleoylphosphatidylcholine (DOPC) doped with two ex trinsic fluorescent probes (NBD-cholesterol/pyrene-PC). Förster re sonance energy transfer (FRET) was observed between the protein and pyre ne-PC and between pyrene-PC and NBD- cholesterol, leading to overlapping emission bands. Partial least squares analysis was applied to fluorescence spectra of pyrene -PC in liposomes with different DOPC/DOPA ratios, generating a model that was tested by an internal validation (leave-one-out cross-validation) and was further used to predict the apparent lipid molar ratio in AChR-containing samples. The values predi cted for DOPA, the lipid with the highest Tm, indicate that the protein exerts a rigidi fying effect on its lipid microenvironment. A similar conclusion was reached from excime r formation of pyrene -PC, a collisional- dependent phenomenon. The excimer/monome r ratio (E/M) at different DOPC/DOPA molar ratios revealed the restricted diffusio n of the probe in AChR-containing samples in comparison to pure lipid samples devoid of protein. FRET from the AChR (donor) to pyrene-PC (acceptor) as a function of temperature was found to increase with increasing temperature, suggesting a shorte r distance between AChR and pyrene PC. Taken together, the results obtained by MA on complex spectra indicate that the AChR rigidifies its surrounding lipid and prefers DOPA rather than DOPC in its immediate microenvironment.
Fil: Wenz, Jorge Javier. 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. Unesco; Argentina
Fil: Barrantes, Francisco Jose. 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. Unesco; Argentina
Materia
Achr
Fluorescencia
Lipido
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/44119
Acceder
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network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Resolution of complex fluorescence spectra of lipids and nicotinic acetylcholine receptor by multivariate analysis reveals protein-mediated effects on the receptor's immediate lipid microenvironmentWenz, Jorge JavierBarrantes, Francisco JoseAchrFluorescenciaLipidohttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Analysis of fluorescent spectra from comp lex biological systems containing various fluorescent probes with overlapping emissi on bands is a challenging task. Valuable information can be extracted from the full spectra, however, by using multivariate analysis (MA) of measurements at differen t wavelengths. We a pplied MA to spectral data of purified Torpedo nicotinic acetylcholine receptor (AChR) protein reconstituted into liposomes made up of diol eoylphosphatidic acid (DOPA) and dioleoylphosphatidylcholine (DOPC) doped with two ex trinsic fluorescent probes (NBD-cholesterol/pyrene-PC). Förster re sonance energy transfer (FRET) was observed between the protein and pyre ne-PC and between pyrene-PC and NBD- cholesterol, leading to overlapping emission bands. Partial least squares analysis was applied to fluorescence spectra of pyrene -PC in liposomes with different DOPC/DOPA ratios, generating a model that was tested by an internal validation (leave-one-out cross-validation) and was further used to predict the apparent lipid molar ratio in AChR-containing samples. The values predi cted for DOPA, the lipid with the highest Tm, indicate that the protein exerts a rigidi fying effect on its lipid microenvironment. A similar conclusion was reached from excime r formation of pyrene -PC, a collisional- dependent phenomenon. The excimer/monome r ratio (E/M) at different DOPC/DOPA molar ratios revealed the restricted diffusio n of the probe in AChR-containing samples in comparison to pure lipid samples devoid of protein. FRET from the AChR (donor) to pyrene-PC (acceptor) as a function of temperature was found to increase with increasing temperature, suggesting a shorte r distance between AChR and pyrene PC. Taken together, the results obtained by MA on complex spectra indicate that the AChR rigidifies its surrounding lipid and prefers DOPA rather than DOPC in its immediate microenvironment.Fil: Wenz, Jorge Javier. 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. Unesco; ArgentinaFil: Barrantes, Francisco Jose. 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. Unesco; ArgentinaSpringer2008-12info: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/11336/44119Wenz, Jorge Javier; Barrantes, Francisco Jose; Resolution of complex fluorescence spectra of lipids and nicotinic acetylcholine receptor by multivariate analysis reveals protein-mediated effects on the receptor's immediate lipid microenvironment; Springer; PMC Biophysics; 1; 1; 12-2008; 1-121757-5036CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://link.springer.com/article/10.1186/1757-5036-1-6info:eu-repo/semantics/altIdentifier/doi/10.1186/1757-5036-1-6info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T14:28:48Zoai:ri.conicet.gov.ar:11336/44119instacron: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:34982025-10-15 14:28:48.693CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Resolution of complex fluorescence spectra of lipids and nicotinic acetylcholine receptor by multivariate analysis reveals protein-mediated effects on the receptor's immediate lipid microenvironment
title Resolution of complex fluorescence spectra of lipids and nicotinic acetylcholine receptor by multivariate analysis reveals protein-mediated effects on the receptor's immediate lipid microenvironment
spellingShingle Resolution of complex fluorescence spectra of lipids and nicotinic acetylcholine receptor by multivariate analysis reveals protein-mediated effects on the receptor's immediate lipid microenvironment
Wenz, Jorge Javier
Achr
Fluorescencia
Lipido
title_short Resolution of complex fluorescence spectra of lipids and nicotinic acetylcholine receptor by multivariate analysis reveals protein-mediated effects on the receptor's immediate lipid microenvironment
title_full Resolution of complex fluorescence spectra of lipids and nicotinic acetylcholine receptor by multivariate analysis reveals protein-mediated effects on the receptor's immediate lipid microenvironment
title_fullStr Resolution of complex fluorescence spectra of lipids and nicotinic acetylcholine receptor by multivariate analysis reveals protein-mediated effects on the receptor's immediate lipid microenvironment
title_full_unstemmed Resolution of complex fluorescence spectra of lipids and nicotinic acetylcholine receptor by multivariate analysis reveals protein-mediated effects on the receptor's immediate lipid microenvironment
title_sort Resolution of complex fluorescence spectra of lipids and nicotinic acetylcholine receptor by multivariate analysis reveals protein-mediated effects on the receptor's immediate lipid microenvironment
dc.creator.none.fl_str_mv Wenz, Jorge Javier
Barrantes, Francisco Jose
author Wenz, Jorge Javier
author_facet Wenz, Jorge Javier
Barrantes, Francisco Jose
author_role author
author2 Barrantes, Francisco Jose
author2_role author
dc.subject.none.fl_str_mv Achr
Fluorescencia
Lipido
topic Achr
Fluorescencia
Lipido
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Analysis of fluorescent spectra from comp lex biological systems containing various fluorescent probes with overlapping emissi on bands is a challenging task. Valuable information can be extracted from the full spectra, however, by using multivariate analysis (MA) of measurements at differen t wavelengths. We a pplied MA to spectral data of purified Torpedo nicotinic acetylcholine receptor (AChR) protein reconstituted into liposomes made up of diol eoylphosphatidic acid (DOPA) and dioleoylphosphatidylcholine (DOPC) doped with two ex trinsic fluorescent probes (NBD-cholesterol/pyrene-PC). Förster re sonance energy transfer (FRET) was observed between the protein and pyre ne-PC and between pyrene-PC and NBD- cholesterol, leading to overlapping emission bands. Partial least squares analysis was applied to fluorescence spectra of pyrene -PC in liposomes with different DOPC/DOPA ratios, generating a model that was tested by an internal validation (leave-one-out cross-validation) and was further used to predict the apparent lipid molar ratio in AChR-containing samples. The values predi cted for DOPA, the lipid with the highest Tm, indicate that the protein exerts a rigidi fying effect on its lipid microenvironment. A similar conclusion was reached from excime r formation of pyrene -PC, a collisional- dependent phenomenon. The excimer/monome r ratio (E/M) at different DOPC/DOPA molar ratios revealed the restricted diffusio n of the probe in AChR-containing samples in comparison to pure lipid samples devoid of protein. FRET from the AChR (donor) to pyrene-PC (acceptor) as a function of temperature was found to increase with increasing temperature, suggesting a shorte r distance between AChR and pyrene PC. Taken together, the results obtained by MA on complex spectra indicate that the AChR rigidifies its surrounding lipid and prefers DOPA rather than DOPC in its immediate microenvironment.
Fil: Wenz, Jorge Javier. 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. Unesco; Argentina
Fil: Barrantes, Francisco Jose. 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. Unesco; Argentina
description Analysis of fluorescent spectra from comp lex biological systems containing various fluorescent probes with overlapping emissi on bands is a challenging task. Valuable information can be extracted from the full spectra, however, by using multivariate analysis (MA) of measurements at differen t wavelengths. We a pplied MA to spectral data of purified Torpedo nicotinic acetylcholine receptor (AChR) protein reconstituted into liposomes made up of diol eoylphosphatidic acid (DOPA) and dioleoylphosphatidylcholine (DOPC) doped with two ex trinsic fluorescent probes (NBD-cholesterol/pyrene-PC). Förster re sonance energy transfer (FRET) was observed between the protein and pyre ne-PC and between pyrene-PC and NBD- cholesterol, leading to overlapping emission bands. Partial least squares analysis was applied to fluorescence spectra of pyrene -PC in liposomes with different DOPC/DOPA ratios, generating a model that was tested by an internal validation (leave-one-out cross-validation) and was further used to predict the apparent lipid molar ratio in AChR-containing samples. The values predi cted for DOPA, the lipid with the highest Tm, indicate that the protein exerts a rigidi fying effect on its lipid microenvironment. A similar conclusion was reached from excime r formation of pyrene -PC, a collisional- dependent phenomenon. The excimer/monome r ratio (E/M) at different DOPC/DOPA molar ratios revealed the restricted diffusio n of the probe in AChR-containing samples in comparison to pure lipid samples devoid of protein. FRET from the AChR (donor) to pyrene-PC (acceptor) as a function of temperature was found to increase with increasing temperature, suggesting a shorte r distance between AChR and pyrene PC. Taken together, the results obtained by MA on complex spectra indicate that the AChR rigidifies its surrounding lipid and prefers DOPA rather than DOPC in its immediate microenvironment.
publishDate 2008
dc.date.none.fl_str_mv 2008-12
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/11336/44119
Wenz, Jorge Javier; Barrantes, Francisco Jose; Resolution of complex fluorescence spectra of lipids and nicotinic acetylcholine receptor by multivariate analysis reveals protein-mediated effects on the receptor's immediate lipid microenvironment; Springer; PMC Biophysics; 1; 1; 12-2008; 1-12
1757-5036
CONICET Digital
CONICET
url http://hdl.handle.net/11336/44119
identifier_str_mv Wenz, Jorge Javier; Barrantes, Francisco Jose; Resolution of complex fluorescence spectra of lipids and nicotinic acetylcholine receptor by multivariate analysis reveals protein-mediated effects on the receptor's immediate lipid microenvironment; Springer; PMC Biophysics; 1; 1; 12-2008; 1-12
1757-5036
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/10.1186/1757-5036-1-6
info:eu-repo/semantics/altIdentifier/doi/10.1186/1757-5036-1-6
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Springer
publisher.none.fl_str_mv Springer
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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