Kinetics of lipid-membrane binding and conformational change of L-BABP

Autores
Galassi, Vanesa Viviana; Nolan, María Verónica; Villarreal, Marcos Ariel; Perducca, Massimiliano; Monaco, Hugo L.; Montich, Guillermo Gabriel
Año de publicación
2009
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
We designed an experimental approach to differentiate the kinetics of protein binding to a lipid membrane from the kinetics of the associated conformational change in the protein. We measured the fluorescence intensity of the single Trp6 in chicken liver bile acid-binding protein (L-BABP) as a function of time after mixing the protein with lipid membranes. We mixed the protein with pure lipid membranes, with lipid membranes in the presence of a soluble quencher, and with lipid membranes containing a fluorescence quencher attached to the lipid polar head group. We fitted simultaneously the experimental curves to a three-state kinetic model. We conclude that in a first step, the binding of L-BABP to the interfacial region of the anionic lipid polar head groups occurred simultaneously with a conformational change to the partly unfolded state. In a second slower step, Trp6 buried within the polar head group region, releasing contacts with the aqueous phase. © 2009 Elsevier Inc. All rights reserved.
Fil: Galassi, Vanesa Viviana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina
Fil: Nolan, María Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina
Fil: Villarreal, Marcos Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina
Fil: Perducca, Massimiliano. Universita di Verona; Italia
Fil: Monaco, Hugo L.. Universita di Verona; Italia
Fil: Montich, Guillermo Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina
Materia
Fluorescence
Kinetics
L-Babp
Protein Binding to Lipid Membrane
Protein Unfolding
Stopped-Flow
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:11336/53929
Acceder
id CONICETDig_00874da4e4045731ed96e75f8f8c7923
oai_identifier_str oai:ri.conicet.gov.ar:11336/53929
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Kinetics of lipid-membrane binding and conformational change of L-BABPGalassi, Vanesa VivianaNolan, María VerónicaVillarreal, Marcos ArielPerducca, MassimilianoMonaco, Hugo L.Montich, Guillermo GabrielFluorescenceKineticsL-BabpProtein Binding to Lipid MembraneProtein UnfoldingStopped-Flowhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1We designed an experimental approach to differentiate the kinetics of protein binding to a lipid membrane from the kinetics of the associated conformational change in the protein. We measured the fluorescence intensity of the single Trp6 in chicken liver bile acid-binding protein (L-BABP) as a function of time after mixing the protein with lipid membranes. We mixed the protein with pure lipid membranes, with lipid membranes in the presence of a soluble quencher, and with lipid membranes containing a fluorescence quencher attached to the lipid polar head group. We fitted simultaneously the experimental curves to a three-state kinetic model. We conclude that in a first step, the binding of L-BABP to the interfacial region of the anionic lipid polar head groups occurred simultaneously with a conformational change to the partly unfolded state. In a second slower step, Trp6 buried within the polar head group region, releasing contacts with the aqueous phase. © 2009 Elsevier Inc. All rights reserved.Fil: Galassi, Vanesa Viviana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Nolan, María Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Villarreal, Marcos Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Perducca, Massimiliano. Universita di Verona; ItaliaFil: Monaco, Hugo L.. Universita di Verona; ItaliaFil: Montich, Guillermo Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; ArgentinaAcademic Press Inc Elsevier Science2009-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/53929Galassi, Vanesa Viviana; Nolan, María Verónica; Villarreal, Marcos Ariel; Perducca, Massimiliano; Monaco, Hugo L.; et al.; Kinetics of lipid-membrane binding and conformational change of L-BABP; Academic Press Inc Elsevier Science; Biochemical and Biophysical Research Communications; 382; 4; 12-2009; 771-7750006-291XCONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0006291X09005956info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bbrc.2009.03.103info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:53:47Zoai:ri.conicet.gov.ar:11336/53929instacron: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:34982025-09-03 09:53:47.707CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Kinetics of lipid-membrane binding and conformational change of L-BABP
title Kinetics of lipid-membrane binding and conformational change of L-BABP
spellingShingle Kinetics of lipid-membrane binding and conformational change of L-BABP
Galassi, Vanesa Viviana
Fluorescence
Kinetics
L-Babp
Protein Binding to Lipid Membrane
Protein Unfolding
Stopped-Flow
title_short Kinetics of lipid-membrane binding and conformational change of L-BABP
title_full Kinetics of lipid-membrane binding and conformational change of L-BABP
title_fullStr Kinetics of lipid-membrane binding and conformational change of L-BABP
title_full_unstemmed Kinetics of lipid-membrane binding and conformational change of L-BABP
title_sort Kinetics of lipid-membrane binding and conformational change of L-BABP
dc.creator.none.fl_str_mv Galassi, Vanesa Viviana
Nolan, María Verónica
Villarreal, Marcos Ariel
Perducca, Massimiliano
Monaco, Hugo L.
Montich, Guillermo Gabriel
author Galassi, Vanesa Viviana
author_facet Galassi, Vanesa Viviana
Nolan, María Verónica
Villarreal, Marcos Ariel
Perducca, Massimiliano
Monaco, Hugo L.
Montich, Guillermo Gabriel
author_role author
author2 Nolan, María Verónica
Villarreal, Marcos Ariel
Perducca, Massimiliano
Monaco, Hugo L.
Montich, Guillermo Gabriel
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Fluorescence
Kinetics
L-Babp
Protein Binding to Lipid Membrane
Protein Unfolding
Stopped-Flow
topic Fluorescence
Kinetics
L-Babp
Protein Binding to Lipid Membrane
Protein Unfolding
Stopped-Flow
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv We designed an experimental approach to differentiate the kinetics of protein binding to a lipid membrane from the kinetics of the associated conformational change in the protein. We measured the fluorescence intensity of the single Trp6 in chicken liver bile acid-binding protein (L-BABP) as a function of time after mixing the protein with lipid membranes. We mixed the protein with pure lipid membranes, with lipid membranes in the presence of a soluble quencher, and with lipid membranes containing a fluorescence quencher attached to the lipid polar head group. We fitted simultaneously the experimental curves to a three-state kinetic model. We conclude that in a first step, the binding of L-BABP to the interfacial region of the anionic lipid polar head groups occurred simultaneously with a conformational change to the partly unfolded state. In a second slower step, Trp6 buried within the polar head group region, releasing contacts with the aqueous phase. © 2009 Elsevier Inc. All rights reserved.
Fil: Galassi, Vanesa Viviana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina
Fil: Nolan, María Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina
Fil: Villarreal, Marcos Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina
Fil: Perducca, Massimiliano. Universita di Verona; Italia
Fil: Monaco, Hugo L.. Universita di Verona; Italia
Fil: Montich, Guillermo Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina
description We designed an experimental approach to differentiate the kinetics of protein binding to a lipid membrane from the kinetics of the associated conformational change in the protein. We measured the fluorescence intensity of the single Trp6 in chicken liver bile acid-binding protein (L-BABP) as a function of time after mixing the protein with lipid membranes. We mixed the protein with pure lipid membranes, with lipid membranes in the presence of a soluble quencher, and with lipid membranes containing a fluorescence quencher attached to the lipid polar head group. We fitted simultaneously the experimental curves to a three-state kinetic model. We conclude that in a first step, the binding of L-BABP to the interfacial region of the anionic lipid polar head groups occurred simultaneously with a conformational change to the partly unfolded state. In a second slower step, Trp6 buried within the polar head group region, releasing contacts with the aqueous phase. © 2009 Elsevier Inc. All rights reserved.
publishDate 2009
dc.date.none.fl_str_mv 2009-12
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/11336/53929
Galassi, Vanesa Viviana; Nolan, María Verónica; Villarreal, Marcos Ariel; Perducca, Massimiliano; Monaco, Hugo L.; et al.; Kinetics of lipid-membrane binding and conformational change of L-BABP; Academic Press Inc Elsevier Science; Biochemical and Biophysical Research Communications; 382; 4; 12-2009; 771-775
0006-291X
CONICET Digital
CONICET
url http://hdl.handle.net/11336/53929
identifier_str_mv Galassi, Vanesa Viviana; Nolan, María Verónica; Villarreal, Marcos Ariel; Perducca, Massimiliano; Monaco, Hugo L.; et al.; Kinetics of lipid-membrane binding and conformational change of L-BABP; Academic Press Inc Elsevier Science; Biochemical and Biophysical Research Communications; 382; 4; 12-2009; 771-775
0006-291X
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://www.sciencedirect.com/science/article/pii/S0006291X09005956
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bbrc.2009.03.103
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
application/pdf
dc.publisher.none.fl_str_mv Academic Press Inc Elsevier Science
publisher.none.fl_str_mv Academic Press Inc Elsevier 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
_version_ 1842269247601901568
score 12.885934

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