Nanoarchitectonic approaches for measuring the catalytic behavior of a membrane anchored enzyme: From Langmuir-Blodgett to a novel Langmuir-Schaefer based nanofilm building device

Autores
Felsztyna, Iván; Perillo, Maria Angelica; Clop, Eduardo Matias
Año de publicación
2023
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Self-organized lipid monolayers at the air-water interface (Langmuir films, LF) are commonly used for measuring the catalytic properties of membrane-bound enzymes. This methodology allows to provide a consistent flat topography molecular density, packing defects and thickness. The aim of the present work was to show the methodological advantages of using the horizontal transfer method (Langmuir-Schaefer) with respect to the vertical transfer method (Langmuir-Blodgett) when mounting a device to measure catalytic activity of membrane enzymes. Based on the results obtained we can conclude that it is possible to prepare stable Langmuir-Blodgett (LB) and Langmuir-Schaefer (LS) films from Bovine Erythrocyte Membranes (BEM) preserving the catalytic activity of its native Acetylcholinesterase (BEA). In comparison, the LS films showed Vmax values more similar to the enzyme present in the vesicles of natural membranes. In addition, it was much easier to produce large amounts of transferred areas with the horizontal transfer methodology. It was possible to decrease the time required to mount an assay with numerous activity points, such as building activity curves as a function of substrate concentration. The present results show that LSBEM provides a proof of concept for the development of biosensors based on transferred purified membrane for the screening of new products acting on an enzyme embedded on its natural milieu. In the case of BEA, the application of these enzymatic sensors could have medical interest, providing drug screening tools for the treatment of Alzheimer's disease.
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: 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
ERYTHROCYTE ACETYLCHOLINESTERASE
ERYTHROCYTE GHOST MEMBRANES
LANGMUIR-BLODGETT FILMS
LANGMUIR-SCHAEFER FILMS
Nivel de accesibilidad
acceso embargado
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:11336/224135
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/224135
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Nanoarchitectonic approaches for measuring the catalytic behavior of a membrane anchored enzyme: From Langmuir-Blodgett to a novel Langmuir-Schaefer based nanofilm building deviceFelsztyna, IvánPerillo, Maria AngelicaClop, Eduardo MatiasBIOSENSOR PROOF OF PRINCIPLEERYTHROCYTE ACETYLCHOLINESTERASEERYTHROCYTE GHOST MEMBRANESLANGMUIR-BLODGETT FILMSLANGMUIR-SCHAEFER FILMShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Self-organized lipid monolayers at the air-water interface (Langmuir films, LF) are commonly used for measuring the catalytic properties of membrane-bound enzymes. This methodology allows to provide a consistent flat topography molecular density, packing defects and thickness. The aim of the present work was to show the methodological advantages of using the horizontal transfer method (Langmuir-Schaefer) with respect to the vertical transfer method (Langmuir-Blodgett) when mounting a device to measure catalytic activity of membrane enzymes. Based on the results obtained we can conclude that it is possible to prepare stable Langmuir-Blodgett (LB) and Langmuir-Schaefer (LS) films from Bovine Erythrocyte Membranes (BEM) preserving the catalytic activity of its native Acetylcholinesterase (BEA). In comparison, the LS films showed Vmax values more similar to the enzyme present in the vesicles of natural membranes. In addition, it was much easier to produce large amounts of transferred areas with the horizontal transfer methodology. It was possible to decrease the time required to mount an assay with numerous activity points, such as building activity curves as a function of substrate concentration. The present results show that LSBEM provides a proof of concept for the development of biosensors based on transferred purified membrane for the screening of new products acting on an enzyme embedded on its natural milieu. In the case of BEA, the application of these enzymatic sensors could have medical interest, providing drug screening tools for the treatment of Alzheimer's disease.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: 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 Science2023-08info:eu-repo/date/embargoEnd/2024-02-18info: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/11336/224135Felsztyna, Iván; Perillo, Maria Angelica; Clop, Eduardo Matias; Nanoarchitectonic approaches for measuring the catalytic behavior of a membrane anchored enzyme: From Langmuir-Blodgett to a novel Langmuir-Schaefer based nanofilm building device; Elsevier Science; Biochimica et Biophysica Acta - Biomembranes; 1865; 6; 8-2023; 1-250005-2736CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://linkinghub.elsevier.com/retrieve/pii/S0005273623000597info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bbamem.2023.184177info:eu-repo/semantics/embargoedAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:56:18Zoai:ri.conicet.gov.ar:11336/224135instacron: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 09:56:18.417CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Nanoarchitectonic approaches for measuring the catalytic behavior of a membrane anchored enzyme: From Langmuir-Blodgett to a novel Langmuir-Schaefer based nanofilm building device
title Nanoarchitectonic approaches for measuring the catalytic behavior of a membrane anchored enzyme: From Langmuir-Blodgett to a novel Langmuir-Schaefer based nanofilm building device
spellingShingle Nanoarchitectonic approaches for measuring the catalytic behavior of a membrane anchored enzyme: From Langmuir-Blodgett to a novel Langmuir-Schaefer based nanofilm building device
Felsztyna, Iván
BIOSENSOR PROOF OF PRINCIPLE
ERYTHROCYTE ACETYLCHOLINESTERASE
ERYTHROCYTE GHOST MEMBRANES
LANGMUIR-BLODGETT FILMS
LANGMUIR-SCHAEFER FILMS
title_short Nanoarchitectonic approaches for measuring the catalytic behavior of a membrane anchored enzyme: From Langmuir-Blodgett to a novel Langmuir-Schaefer based nanofilm building device
title_full Nanoarchitectonic approaches for measuring the catalytic behavior of a membrane anchored enzyme: From Langmuir-Blodgett to a novel Langmuir-Schaefer based nanofilm building device
title_fullStr Nanoarchitectonic approaches for measuring the catalytic behavior of a membrane anchored enzyme: From Langmuir-Blodgett to a novel Langmuir-Schaefer based nanofilm building device
title_full_unstemmed Nanoarchitectonic approaches for measuring the catalytic behavior of a membrane anchored enzyme: From Langmuir-Blodgett to a novel Langmuir-Schaefer based nanofilm building device
title_sort Nanoarchitectonic approaches for measuring the catalytic behavior of a membrane anchored enzyme: From Langmuir-Blodgett to a novel Langmuir-Schaefer based nanofilm building device
dc.creator.none.fl_str_mv Felsztyna, Iván
Perillo, Maria Angelica
Clop, Eduardo Matias
author Felsztyna, Iván
author_facet Felsztyna, Iván
Perillo, Maria Angelica
Clop, Eduardo Matias
author_role author
author2 Perillo, Maria Angelica
Clop, Eduardo Matias
author2_role author
author
dc.subject.none.fl_str_mv BIOSENSOR PROOF OF PRINCIPLE
ERYTHROCYTE ACETYLCHOLINESTERASE
ERYTHROCYTE GHOST MEMBRANES
LANGMUIR-BLODGETT FILMS
LANGMUIR-SCHAEFER FILMS
topic BIOSENSOR PROOF OF PRINCIPLE
ERYTHROCYTE ACETYLCHOLINESTERASE
ERYTHROCYTE GHOST MEMBRANES
LANGMUIR-BLODGETT FILMS
LANGMUIR-SCHAEFER 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 Self-organized lipid monolayers at the air-water interface (Langmuir films, LF) are commonly used for measuring the catalytic properties of membrane-bound enzymes. This methodology allows to provide a consistent flat topography molecular density, packing defects and thickness. The aim of the present work was to show the methodological advantages of using the horizontal transfer method (Langmuir-Schaefer) with respect to the vertical transfer method (Langmuir-Blodgett) when mounting a device to measure catalytic activity of membrane enzymes. Based on the results obtained we can conclude that it is possible to prepare stable Langmuir-Blodgett (LB) and Langmuir-Schaefer (LS) films from Bovine Erythrocyte Membranes (BEM) preserving the catalytic activity of its native Acetylcholinesterase (BEA). In comparison, the LS films showed Vmax values more similar to the enzyme present in the vesicles of natural membranes. In addition, it was much easier to produce large amounts of transferred areas with the horizontal transfer methodology. It was possible to decrease the time required to mount an assay with numerous activity points, such as building activity curves as a function of substrate concentration. The present results show that LSBEM provides a proof of concept for the development of biosensors based on transferred purified membrane for the screening of new products acting on an enzyme embedded on its natural milieu. In the case of BEA, the application of these enzymatic sensors could have medical interest, providing drug screening tools for the treatment of Alzheimer's disease.
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: 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 Self-organized lipid monolayers at the air-water interface (Langmuir films, LF) are commonly used for measuring the catalytic properties of membrane-bound enzymes. This methodology allows to provide a consistent flat topography molecular density, packing defects and thickness. The aim of the present work was to show the methodological advantages of using the horizontal transfer method (Langmuir-Schaefer) with respect to the vertical transfer method (Langmuir-Blodgett) when mounting a device to measure catalytic activity of membrane enzymes. Based on the results obtained we can conclude that it is possible to prepare stable Langmuir-Blodgett (LB) and Langmuir-Schaefer (LS) films from Bovine Erythrocyte Membranes (BEM) preserving the catalytic activity of its native Acetylcholinesterase (BEA). In comparison, the LS films showed Vmax values more similar to the enzyme present in the vesicles of natural membranes. In addition, it was much easier to produce large amounts of transferred areas with the horizontal transfer methodology. It was possible to decrease the time required to mount an assay with numerous activity points, such as building activity curves as a function of substrate concentration. The present results show that LSBEM provides a proof of concept for the development of biosensors based on transferred purified membrane for the screening of new products acting on an enzyme embedded on its natural milieu. In the case of BEA, the application of these enzymatic sensors could have medical interest, providing drug screening tools for the treatment of Alzheimer's disease.
publishDate 2023
dc.date.none.fl_str_mv 2023-08
info:eu-repo/date/embargoEnd/2024-02-18
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/11336/224135
Felsztyna, Iván; Perillo, Maria Angelica; Clop, Eduardo Matias; Nanoarchitectonic approaches for measuring the catalytic behavior of a membrane anchored enzyme: From Langmuir-Blodgett to a novel Langmuir-Schaefer based nanofilm building device; Elsevier Science; Biochimica et Biophysica Acta - Biomembranes; 1865; 6; 8-2023; 1-25
0005-2736
CONICET Digital
CONICET
url http://hdl.handle.net/11336/224135
identifier_str_mv Felsztyna, Iván; Perillo, Maria Angelica; Clop, Eduardo Matias; Nanoarchitectonic approaches for measuring the catalytic behavior of a membrane anchored enzyme: From Langmuir-Blodgett to a novel Langmuir-Schaefer based nanofilm building device; Elsevier Science; Biochimica et Biophysica Acta - Biomembranes; 1865; 6; 8-2023; 1-25
0005-2736
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://linkinghub.elsevier.com/retrieve/pii/S0005273623000597
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bbamem.2023.184177
dc.rights.none.fl_str_mv info:eu-repo/semantics/embargoedAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv embargoedAccess
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
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)
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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