A constant area monolayer method to assess optimal lipid packing for lipolysis tested with several secreted phospholipases A2

Autores
Yunes Quartino, Pablo Javier; Portela, Madelón; Lima, Analia Ethel; Duran, Rosario; Lomonte, Bruno; Fidelio, Gerardo Daniel
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
We present an analysis of lipid monolayer hydrolysis at a constant area to assess the optimal lateral surface pressure value (Πopt) and thus, the surface packing density of the lipid, at which the activity of a given lipolytic enzyme is maximal. This isochoric method consists of a measurement of the decrease down to zero of the Πopt of phospholipid substrate monolayer due to continuous hydrolysis using only one reaction compartment. We performed the comparison of both approaches using several commercially available and literature-evaluated sPLA2s. Also, we characterized for the first time the profile of hydrolysis of DLPC monolayers catalyzed by a sPLA2 from Streptomyces violaceoruber and isoenzymes purified from Bothrops diporus venom. One of these viper venom enzymes is a new isoenzyme, partially sequenced by a mass spectrometry approach. We also included the basic myotoxin sPLA2-III from Bothrops asper. Results obtained with the isochoric method and the standard isobaric one produced quite similar values of Πopt, validating the proposal. In addition, we propose a new classification parameter, a lipolytic ratio of hydrolysis at two lateral pressures, 20 mN·m− 1 and 10 mN·m− 1, termed here as LR20/10 index. This index differentiates quite well “high surface pressure” from “low surface pressure” sPLA2s and, by extension; it can be used as a functional criterion for the quality of a certain enzyme. Also, this index could be added to the grouping systematic criteria for the superfamily proposed for phospholipase A2.
Fil: Yunes Quartino, Pablo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Ciencia y Tecnología de Alimentos Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Ciencia y Tecnología de Alimentos Córdoba; Argentina
Fil: Portela, Madelón. Institut Pasteur de Montevideo. Instituto de Investigaciones Biológicas Clemente Estable, Unidad de Bioquímica y Proteómica Analíticas; Uruguay
Fil: Lima, Analia Ethel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Duran, Rosario. Institut Pasteur de Montevideo. Instituto de Investigaciones Biológicas Clemente Estable, Unidad de Bioquímica y Proteómica Analíticas; Uruguay
Fil: Lomonte, Bruno. Universidad de Costa Rica; Costa Rica
Fil: Fidelio, Gerardo Daniel. 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
Lipid Monolayer
Phospholipase A2
Surface Preassure
Isochoric Method
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:11336/50731
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/50731
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling A constant area monolayer method to assess optimal lipid packing for lipolysis tested with several secreted phospholipases A2Yunes Quartino, Pablo JavierPortela, MadelónLima, Analia EthelDuran, RosarioLomonte, BrunoFidelio, Gerardo DanielLipid MonolayerPhospholipase A2Surface PreassureIsochoric Methodhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1We present an analysis of lipid monolayer hydrolysis at a constant area to assess the optimal lateral surface pressure value (Πopt) and thus, the surface packing density of the lipid, at which the activity of a given lipolytic enzyme is maximal. This isochoric method consists of a measurement of the decrease down to zero of the Πopt of phospholipid substrate monolayer due to continuous hydrolysis using only one reaction compartment. We performed the comparison of both approaches using several commercially available and literature-evaluated sPLA2s. Also, we characterized for the first time the profile of hydrolysis of DLPC monolayers catalyzed by a sPLA2 from Streptomyces violaceoruber and isoenzymes purified from Bothrops diporus venom. One of these viper venom enzymes is a new isoenzyme, partially sequenced by a mass spectrometry approach. We also included the basic myotoxin sPLA2-III from Bothrops asper. Results obtained with the isochoric method and the standard isobaric one produced quite similar values of Πopt, validating the proposal. In addition, we propose a new classification parameter, a lipolytic ratio of hydrolysis at two lateral pressures, 20 mN·m− 1 and 10 mN·m− 1, termed here as LR20/10 index. This index differentiates quite well “high surface pressure” from “low surface pressure” sPLA2s and, by extension; it can be used as a functional criterion for the quality of a certain enzyme. Also, this index could be added to the grouping systematic criteria for the superfamily proposed for phospholipase A2.Fil: Yunes Quartino, Pablo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Ciencia y Tecnología de Alimentos Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Ciencia y Tecnología de Alimentos Córdoba; ArgentinaFil: Portela, Madelón. Institut Pasteur de Montevideo. Instituto de Investigaciones Biológicas Clemente Estable, Unidad de Bioquímica y Proteómica Analíticas; UruguayFil: Lima, Analia Ethel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Duran, Rosario. Institut Pasteur de Montevideo. Instituto de Investigaciones Biológicas Clemente Estable, Unidad de Bioquímica y Proteómica Analíticas; UruguayFil: Lomonte, Bruno. Universidad de Costa Rica; Costa RicaFil: Fidelio, Gerardo Daniel. 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; ArgentinaElsevier Science2015-10info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/50731Yunes Quartino, Pablo Javier; Portela, Madelón; Lima, Analia Ethel; Duran, Rosario; Lomonte, Bruno; et al.; A constant area monolayer method to assess optimal lipid packing for lipolysis tested with several secreted phospholipases A2; Elsevier Science; Biochimica et Biophysica Acta - Biomembranes; 1848; 10; 10-2015; 2216-22240005-2736CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0005273615001819info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bbamem.2015.06.003info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:54:25Zoai:ri.conicet.gov.ar:11336/50731instacron: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 09:54:25.426CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv A constant area monolayer method to assess optimal lipid packing for lipolysis tested with several secreted phospholipases A2
title A constant area monolayer method to assess optimal lipid packing for lipolysis tested with several secreted phospholipases A2
spellingShingle A constant area monolayer method to assess optimal lipid packing for lipolysis tested with several secreted phospholipases A2
Yunes Quartino, Pablo Javier
Lipid Monolayer
Phospholipase A2
Surface Preassure
Isochoric Method
title_short A constant area monolayer method to assess optimal lipid packing for lipolysis tested with several secreted phospholipases A2
title_full A constant area monolayer method to assess optimal lipid packing for lipolysis tested with several secreted phospholipases A2
title_fullStr A constant area monolayer method to assess optimal lipid packing for lipolysis tested with several secreted phospholipases A2
title_full_unstemmed A constant area monolayer method to assess optimal lipid packing for lipolysis tested with several secreted phospholipases A2
title_sort A constant area monolayer method to assess optimal lipid packing for lipolysis tested with several secreted phospholipases A2
dc.creator.none.fl_str_mv Yunes Quartino, Pablo Javier
Portela, Madelón
Lima, Analia Ethel
Duran, Rosario
Lomonte, Bruno
Fidelio, Gerardo Daniel
author Yunes Quartino, Pablo Javier
author_facet Yunes Quartino, Pablo Javier
Portela, Madelón
Lima, Analia Ethel
Duran, Rosario
Lomonte, Bruno
Fidelio, Gerardo Daniel
author_role author
author2 Portela, Madelón
Lima, Analia Ethel
Duran, Rosario
Lomonte, Bruno
Fidelio, Gerardo Daniel
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Lipid Monolayer
Phospholipase A2
Surface Preassure
Isochoric Method
topic Lipid Monolayer
Phospholipase A2
Surface Preassure
Isochoric Method
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 present an analysis of lipid monolayer hydrolysis at a constant area to assess the optimal lateral surface pressure value (Πopt) and thus, the surface packing density of the lipid, at which the activity of a given lipolytic enzyme is maximal. This isochoric method consists of a measurement of the decrease down to zero of the Πopt of phospholipid substrate monolayer due to continuous hydrolysis using only one reaction compartment. We performed the comparison of both approaches using several commercially available and literature-evaluated sPLA2s. Also, we characterized for the first time the profile of hydrolysis of DLPC monolayers catalyzed by a sPLA2 from Streptomyces violaceoruber and isoenzymes purified from Bothrops diporus venom. One of these viper venom enzymes is a new isoenzyme, partially sequenced by a mass spectrometry approach. We also included the basic myotoxin sPLA2-III from Bothrops asper. Results obtained with the isochoric method and the standard isobaric one produced quite similar values of Πopt, validating the proposal. In addition, we propose a new classification parameter, a lipolytic ratio of hydrolysis at two lateral pressures, 20 mN·m− 1 and 10 mN·m− 1, termed here as LR20/10 index. This index differentiates quite well “high surface pressure” from “low surface pressure” sPLA2s and, by extension; it can be used as a functional criterion for the quality of a certain enzyme. Also, this index could be added to the grouping systematic criteria for the superfamily proposed for phospholipase A2.
Fil: Yunes Quartino, Pablo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Ciencia y Tecnología de Alimentos Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Ciencia y Tecnología de Alimentos Córdoba; Argentina
Fil: Portela, Madelón. Institut Pasteur de Montevideo. Instituto de Investigaciones Biológicas Clemente Estable, Unidad de Bioquímica y Proteómica Analíticas; Uruguay
Fil: Lima, Analia Ethel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Duran, Rosario. Institut Pasteur de Montevideo. Instituto de Investigaciones Biológicas Clemente Estable, Unidad de Bioquímica y Proteómica Analíticas; Uruguay
Fil: Lomonte, Bruno. Universidad de Costa Rica; Costa Rica
Fil: Fidelio, Gerardo Daniel. 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 present an analysis of lipid monolayer hydrolysis at a constant area to assess the optimal lateral surface pressure value (Πopt) and thus, the surface packing density of the lipid, at which the activity of a given lipolytic enzyme is maximal. This isochoric method consists of a measurement of the decrease down to zero of the Πopt of phospholipid substrate monolayer due to continuous hydrolysis using only one reaction compartment. We performed the comparison of both approaches using several commercially available and literature-evaluated sPLA2s. Also, we characterized for the first time the profile of hydrolysis of DLPC monolayers catalyzed by a sPLA2 from Streptomyces violaceoruber and isoenzymes purified from Bothrops diporus venom. One of these viper venom enzymes is a new isoenzyme, partially sequenced by a mass spectrometry approach. We also included the basic myotoxin sPLA2-III from Bothrops asper. Results obtained with the isochoric method and the standard isobaric one produced quite similar values of Πopt, validating the proposal. In addition, we propose a new classification parameter, a lipolytic ratio of hydrolysis at two lateral pressures, 20 mN·m− 1 and 10 mN·m− 1, termed here as LR20/10 index. This index differentiates quite well “high surface pressure” from “low surface pressure” sPLA2s and, by extension; it can be used as a functional criterion for the quality of a certain enzyme. Also, this index could be added to the grouping systematic criteria for the superfamily proposed for phospholipase A2.
publishDate 2015
dc.date.none.fl_str_mv 2015-10
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/11336/50731
Yunes Quartino, Pablo Javier; Portela, Madelón; Lima, Analia Ethel; Duran, Rosario; Lomonte, Bruno; et al.; A constant area monolayer method to assess optimal lipid packing for lipolysis tested with several secreted phospholipases A2; Elsevier Science; Biochimica et Biophysica Acta - Biomembranes; 1848; 10; 10-2015; 2216-2224
0005-2736
CONICET Digital
CONICET
url http://hdl.handle.net/11336/50731
identifier_str_mv Yunes Quartino, Pablo Javier; Portela, Madelón; Lima, Analia Ethel; Duran, Rosario; Lomonte, Bruno; et al.; A constant area monolayer method to assess optimal lipid packing for lipolysis tested with several secreted phospholipases A2; Elsevier Science; Biochimica et Biophysica Acta - Biomembranes; 1848; 10; 10-2015; 2216-2224
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://www.sciencedirect.com/science/article/pii/S0005273615001819
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bbamem.2015.06.003
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 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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score 13.13397

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