Sensing molecular organizational changes through the catalytic activity of acetylcholinesterase from erythrocyte membranes in Langmuir-Blodgett films

Autores
Felsztyna, Iván; Turina, Anahi del Valle; Perillo, Maria Angelica; Clop, Eduardo Matias
Año de publicación
2020
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Langmuir films prepared from bovine erythrocyte membranes (LFBEM) were studied and transferred to alkylated glasses (Langmuir-Blodgett films, LBBEM) in order to assess the effects of membrane molecular packing on Bovine Erythrocyte Acetylcholinesterase (BEA) catalytic activity. Surface pressure (π) vs Area isotherms showed three 2D-transitions at ~7, ~18 and ~44 mN/m and a collapse pressure at πc = .49 mN/m. The 0?12?0 mN/m compression-decompression cycles resulted reversible while those 0?40?0 mN/m exhibited a significant hysteresis. Taken together, EFM, BAM and AFM images and the stability of the film after 3C-D cycles, we can suggest that over the air-water interface as well as over the silanized glass substrate the surface is mostly covered by a monolayer with a few particles dispersed. Acetylthiocholine hydrolysis was assayed with BEA in bovine erythrocyte membrane suspensions (SBEM) and in LBBEM packed at 10 (LBBEM,10) and 35 mN/m (LBBEM,35), which gave the following kinetic parameters: Vmax = .3.41 ± 0.15, 0.021 ± 0.002 and 0.030 ± 0.003 nmol.min− 1·μg prot− 1 and KM = .0.11 ± 0.02, 0.047 ± 0.017 and 0.026 ± 0.017 mM, respectively. Although from SBEM to LBBEM we lost active enzyme, the catalytic efficiency (Vmax/KM) increased ~750 times. Eugenol and 1,8-cineol inhibited BEA catalytic activity in LBBEM,35. Our results demonstrate the transmission of information between the membrane and the environment within the subphase immediately below the membrane, where anchored proteins are hosted. This was reflected by the membrane packing-induced modulation of BEA catalytic activity. Furthermore, LBBEM provides a proof of concept for the development of biosensors to screen new green pesticides acting through BEA interaction.
Fil: Felsztyna, Iván. 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: 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: 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
Fil: Clop, Eduardo Matias. 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
BIOSENSOR PROOF OF PRINCIPLE
BOVINE ERYTHROCYTE MEMBRANES
ERYTHROCYTE ACETYLCHOLINESTERASE
LANGMUIR-BLODGETT FILMS
MONOTERPENES
Nivel de accesibilidad
acceso embargado
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/105375
Acceder
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network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Sensing molecular organizational changes through the catalytic activity of acetylcholinesterase from erythrocyte membranes in Langmuir-Blodgett filmsFelsztyna, IvánTurina, Anahi del VallePerillo, Maria AngelicaClop, Eduardo MatiasBIOSENSOR PROOF OF PRINCIPLEBOVINE ERYTHROCYTE MEMBRANESERYTHROCYTE ACETYLCHOLINESTERASELANGMUIR-BLODGETT FILMSMONOTERPENEShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Langmuir films prepared from bovine erythrocyte membranes (LFBEM) were studied and transferred to alkylated glasses (Langmuir-Blodgett films, LBBEM) in order to assess the effects of membrane molecular packing on Bovine Erythrocyte Acetylcholinesterase (BEA) catalytic activity. Surface pressure (π) vs Area isotherms showed three 2D-transitions at ~7, ~18 and ~44 mN/m and a collapse pressure at πc = .49 mN/m. The 0?12?0 mN/m compression-decompression cycles resulted reversible while those 0?40?0 mN/m exhibited a significant hysteresis. Taken together, EFM, BAM and AFM images and the stability of the film after 3C-D cycles, we can suggest that over the air-water interface as well as over the silanized glass substrate the surface is mostly covered by a monolayer with a few particles dispersed. Acetylthiocholine hydrolysis was assayed with BEA in bovine erythrocyte membrane suspensions (SBEM) and in LBBEM packed at 10 (LBBEM,10) and 35 mN/m (LBBEM,35), which gave the following kinetic parameters: Vmax = .3.41 ± 0.15, 0.021 ± 0.002 and 0.030 ± 0.003 nmol.min− 1·μg prot− 1 and KM = .0.11 ± 0.02, 0.047 ± 0.017 and 0.026 ± 0.017 mM, respectively. Although from SBEM to LBBEM we lost active enzyme, the catalytic efficiency (Vmax/KM) increased ~750 times. Eugenol and 1,8-cineol inhibited BEA catalytic activity in LBBEM,35. Our results demonstrate the transmission of information between the membrane and the environment within the subphase immediately below the membrane, where anchored proteins are hosted. This was reflected by the membrane packing-induced modulation of BEA catalytic activity. Furthermore, LBBEM provides a proof of concept for the development of biosensors to screen new green pesticides acting through BEA interaction.Fil: Felsztyna, Iván. 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: 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: 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; ArgentinaFil: Clop, Eduardo Matias. 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; ArgentinaElsevier Science2020-05-01info:eu-repo/date/embargoEnd/2021-05-01info: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/11336/105375Felsztyna, Iván; Turina, Anahi del Valle; Perillo, Maria Angelica; Clop, Eduardo Matias; Sensing molecular organizational changes through the catalytic activity of acetylcholinesterase from erythrocyte membranes in Langmuir-Blodgett films; Elsevier Science; Biochimica et Biophysica Acta - Biomembranes; 1862; 5; 1-5-2020; e183188-e1831880005-27361879-2642CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/abs/pii/S0005273620300146info:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pubmed/31930963info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bbamem.2020.183188info:eu-repo/semantics/embargoedAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:09:33Zoai:ri.conicet.gov.ar:11336/105375instacron: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 10:09:33.392CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Sensing molecular organizational changes through the catalytic activity of acetylcholinesterase from erythrocyte membranes in Langmuir-Blodgett films
title Sensing molecular organizational changes through the catalytic activity of acetylcholinesterase from erythrocyte membranes in Langmuir-Blodgett films
spellingShingle Sensing molecular organizational changes through the catalytic activity of acetylcholinesterase from erythrocyte membranes in Langmuir-Blodgett films
Felsztyna, Iván
BIOSENSOR PROOF OF PRINCIPLE
BOVINE ERYTHROCYTE MEMBRANES
ERYTHROCYTE ACETYLCHOLINESTERASE
LANGMUIR-BLODGETT FILMS
MONOTERPENES
title_short Sensing molecular organizational changes through the catalytic activity of acetylcholinesterase from erythrocyte membranes in Langmuir-Blodgett films
title_full Sensing molecular organizational changes through the catalytic activity of acetylcholinesterase from erythrocyte membranes in Langmuir-Blodgett films
title_fullStr Sensing molecular organizational changes through the catalytic activity of acetylcholinesterase from erythrocyte membranes in Langmuir-Blodgett films
title_full_unstemmed Sensing molecular organizational changes through the catalytic activity of acetylcholinesterase from erythrocyte membranes in Langmuir-Blodgett films
title_sort Sensing molecular organizational changes through the catalytic activity of acetylcholinesterase from erythrocyte membranes in Langmuir-Blodgett films
dc.creator.none.fl_str_mv Felsztyna, Iván
Turina, Anahi del Valle
Perillo, Maria Angelica
Clop, Eduardo Matias
author Felsztyna, Iván
author_facet Felsztyna, Iván
Turina, Anahi del Valle
Perillo, Maria Angelica
Clop, Eduardo Matias
author_role author
author2 Turina, Anahi del Valle
Perillo, Maria Angelica
Clop, Eduardo Matias
author2_role author
author
author
dc.subject.none.fl_str_mv BIOSENSOR PROOF OF PRINCIPLE
BOVINE ERYTHROCYTE MEMBRANES
ERYTHROCYTE ACETYLCHOLINESTERASE
LANGMUIR-BLODGETT FILMS
MONOTERPENES
topic BIOSENSOR PROOF OF PRINCIPLE
BOVINE ERYTHROCYTE MEMBRANES
ERYTHROCYTE ACETYLCHOLINESTERASE
LANGMUIR-BLODGETT FILMS
MONOTERPENES
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Langmuir films prepared from bovine erythrocyte membranes (LFBEM) were studied and transferred to alkylated glasses (Langmuir-Blodgett films, LBBEM) in order to assess the effects of membrane molecular packing on Bovine Erythrocyte Acetylcholinesterase (BEA) catalytic activity. Surface pressure (π) vs Area isotherms showed three 2D-transitions at ~7, ~18 and ~44 mN/m and a collapse pressure at πc = .49 mN/m. The 0?12?0 mN/m compression-decompression cycles resulted reversible while those 0?40?0 mN/m exhibited a significant hysteresis. Taken together, EFM, BAM and AFM images and the stability of the film after 3C-D cycles, we can suggest that over the air-water interface as well as over the silanized glass substrate the surface is mostly covered by a monolayer with a few particles dispersed. Acetylthiocholine hydrolysis was assayed with BEA in bovine erythrocyte membrane suspensions (SBEM) and in LBBEM packed at 10 (LBBEM,10) and 35 mN/m (LBBEM,35), which gave the following kinetic parameters: Vmax = .3.41 ± 0.15, 0.021 ± 0.002 and 0.030 ± 0.003 nmol.min− 1·μg prot− 1 and KM = .0.11 ± 0.02, 0.047 ± 0.017 and 0.026 ± 0.017 mM, respectively. Although from SBEM to LBBEM we lost active enzyme, the catalytic efficiency (Vmax/KM) increased ~750 times. Eugenol and 1,8-cineol inhibited BEA catalytic activity in LBBEM,35. Our results demonstrate the transmission of information between the membrane and the environment within the subphase immediately below the membrane, where anchored proteins are hosted. This was reflected by the membrane packing-induced modulation of BEA catalytic activity. Furthermore, LBBEM provides a proof of concept for the development of biosensors to screen new green pesticides acting through BEA interaction.
Fil: Felsztyna, Iván. 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: 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: 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
Fil: Clop, Eduardo Matias. 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 Langmuir films prepared from bovine erythrocyte membranes (LFBEM) were studied and transferred to alkylated glasses (Langmuir-Blodgett films, LBBEM) in order to assess the effects of membrane molecular packing on Bovine Erythrocyte Acetylcholinesterase (BEA) catalytic activity. Surface pressure (π) vs Area isotherms showed three 2D-transitions at ~7, ~18 and ~44 mN/m and a collapse pressure at πc = .49 mN/m. The 0?12?0 mN/m compression-decompression cycles resulted reversible while those 0?40?0 mN/m exhibited a significant hysteresis. Taken together, EFM, BAM and AFM images and the stability of the film after 3C-D cycles, we can suggest that over the air-water interface as well as over the silanized glass substrate the surface is mostly covered by a monolayer with a few particles dispersed. Acetylthiocholine hydrolysis was assayed with BEA in bovine erythrocyte membrane suspensions (SBEM) and in LBBEM packed at 10 (LBBEM,10) and 35 mN/m (LBBEM,35), which gave the following kinetic parameters: Vmax = .3.41 ± 0.15, 0.021 ± 0.002 and 0.030 ± 0.003 nmol.min− 1·μg prot− 1 and KM = .0.11 ± 0.02, 0.047 ± 0.017 and 0.026 ± 0.017 mM, respectively. Although from SBEM to LBBEM we lost active enzyme, the catalytic efficiency (Vmax/KM) increased ~750 times. Eugenol and 1,8-cineol inhibited BEA catalytic activity in LBBEM,35. Our results demonstrate the transmission of information between the membrane and the environment within the subphase immediately below the membrane, where anchored proteins are hosted. This was reflected by the membrane packing-induced modulation of BEA catalytic activity. Furthermore, LBBEM provides a proof of concept for the development of biosensors to screen new green pesticides acting through BEA interaction.
publishDate 2020
dc.date.none.fl_str_mv 2020-05-01
info:eu-repo/date/embargoEnd/2021-05-01
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/11336/105375
Felsztyna, Iván; Turina, Anahi del Valle; Perillo, Maria Angelica; Clop, Eduardo Matias; Sensing molecular organizational changes through the catalytic activity of acetylcholinesterase from erythrocyte membranes in Langmuir-Blodgett films; Elsevier Science; Biochimica et Biophysica Acta - Biomembranes; 1862; 5; 1-5-2020; e183188-e183188
0005-2736
1879-2642
CONICET Digital
CONICET
url http://hdl.handle.net/11336/105375
identifier_str_mv Felsztyna, Iván; Turina, Anahi del Valle; Perillo, Maria Angelica; Clop, Eduardo Matias; Sensing molecular organizational changes through the catalytic activity of acetylcholinesterase from erythrocyte membranes in Langmuir-Blodgett films; Elsevier Science; Biochimica et Biophysica Acta - Biomembranes; 1862; 5; 1-5-2020; e183188-e183188
0005-2736
1879-2642
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/abs/pii/S0005273620300146
info:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pubmed/31930963
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bbamem.2020.183188
dc.rights.none.fl_str_mv info:eu-repo/semantics/embargoedAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv embargoedAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
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dc.publisher.none.fl_str_mv Elsevier Science
publisher.none.fl_str_mv Elsevier Science
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
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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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