Cloning and functional expression of secreted phospholipases A2 from Bothrops diporus (Yarará Chica)

Autores
Yunes Quartino, Pablo Javier; Barra, Jose Luis; Fidelio, Gerardo Daniel
Año de publicación
2012
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Bothrops diporus is a very common viper in Argentina. At present, no complete sequence of secreted phospholipase A2 (sPLA2) from this snake has been reported. We have cloned two sPLA2 isoenzymes as well as a putative sPLA2-like myotoxin from venom gland. The two sPLA2 were expressed as inclusion bodies in Escherichia coli with an N-terminal tag of ubiquitin. After in vitro renaturation and cleavage step, using an ubiquitin specific peptidase, the recombinants exhibited sPLA2 activity when analyzed by means of Langmuir dilauroylphosphatidylcholine monolayers as substrate. Both enzymes have a similar surface pressure-activity profile when compared with non-recombinant purified isoforms. To our knowledge, this is the first time that analysis of optimal lateral pressure of substrate monolayers by using the surface barostat technique is performed on recombinant sPLA2s.
Fil: Yunes Quartino, Pablo Javier. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Química Biológica; Argentina. 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: Barra, Jose Luis. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Química Biológica; Argentina. 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
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. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Química Biológica; Argentina
Materia
BOTHROPS DIPORUS
DLPC MONOLAYER
PHOSPHOLIPASE A2
SNAKE VENOM
UBIQUITIN
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:11336/198118
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/198118
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Cloning and functional expression of secreted phospholipases A2 from Bothrops diporus (Yarará Chica)Yunes Quartino, Pablo JavierBarra, Jose LuisFidelio, Gerardo DanielBOTHROPS DIPORUSDLPC MONOLAYERPHOSPHOLIPASE A2SNAKE VENOMUBIQUITINhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Bothrops diporus is a very common viper in Argentina. At present, no complete sequence of secreted phospholipase A2 (sPLA2) from this snake has been reported. We have cloned two sPLA2 isoenzymes as well as a putative sPLA2-like myotoxin from venom gland. The two sPLA2 were expressed as inclusion bodies in Escherichia coli with an N-terminal tag of ubiquitin. After in vitro renaturation and cleavage step, using an ubiquitin specific peptidase, the recombinants exhibited sPLA2 activity when analyzed by means of Langmuir dilauroylphosphatidylcholine monolayers as substrate. Both enzymes have a similar surface pressure-activity profile when compared with non-recombinant purified isoforms. To our knowledge, this is the first time that analysis of optimal lateral pressure of substrate monolayers by using the surface barostat technique is performed on recombinant sPLA2s.Fil: Yunes Quartino, Pablo Javier. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Química Biológica; Argentina. 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: Barra, Jose Luis. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Química Biológica; Argentina. 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; ArgentinaFil: 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. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Química Biológica; ArgentinaAcademic Press Inc Elsevier Science2012-10info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/198118Yunes Quartino, Pablo Javier; Barra, Jose Luis; Fidelio, Gerardo Daniel; Cloning and functional expression of secreted phospholipases A2 from Bothrops diporus (Yarará Chica); Academic Press Inc Elsevier Science; Biochemical and Biophysical Research Communications; 427; 2; 10-2012; 321-3250006-291XCONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0006291X12017913info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bbrc.2012.09.051info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-10T13:22:18Zoai:ri.conicet.gov.ar:11336/198118instacron: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:34982025-09-10 13:22:18.85CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Cloning and functional expression of secreted phospholipases A2 from Bothrops diporus (Yarará Chica)
title Cloning and functional expression of secreted phospholipases A2 from Bothrops diporus (Yarará Chica)
spellingShingle Cloning and functional expression of secreted phospholipases A2 from Bothrops diporus (Yarará Chica)
Yunes Quartino, Pablo Javier
BOTHROPS DIPORUS
DLPC MONOLAYER
PHOSPHOLIPASE A2
SNAKE VENOM
UBIQUITIN
title_short Cloning and functional expression of secreted phospholipases A2 from Bothrops diporus (Yarará Chica)
title_full Cloning and functional expression of secreted phospholipases A2 from Bothrops diporus (Yarará Chica)
title_fullStr Cloning and functional expression of secreted phospholipases A2 from Bothrops diporus (Yarará Chica)
title_full_unstemmed Cloning and functional expression of secreted phospholipases A2 from Bothrops diporus (Yarará Chica)
title_sort Cloning and functional expression of secreted phospholipases A2 from Bothrops diporus (Yarará Chica)
dc.creator.none.fl_str_mv Yunes Quartino, Pablo Javier
Barra, Jose Luis
Fidelio, Gerardo Daniel
author Yunes Quartino, Pablo Javier
author_facet Yunes Quartino, Pablo Javier
Barra, Jose Luis
Fidelio, Gerardo Daniel
author_role author
author2 Barra, Jose Luis
Fidelio, Gerardo Daniel
author2_role author
author
dc.subject.none.fl_str_mv BOTHROPS DIPORUS
DLPC MONOLAYER
PHOSPHOLIPASE A2
SNAKE VENOM
UBIQUITIN
topic BOTHROPS DIPORUS
DLPC MONOLAYER
PHOSPHOLIPASE A2
SNAKE VENOM
UBIQUITIN
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Bothrops diporus is a very common viper in Argentina. At present, no complete sequence of secreted phospholipase A2 (sPLA2) from this snake has been reported. We have cloned two sPLA2 isoenzymes as well as a putative sPLA2-like myotoxin from venom gland. The two sPLA2 were expressed as inclusion bodies in Escherichia coli with an N-terminal tag of ubiquitin. After in vitro renaturation and cleavage step, using an ubiquitin specific peptidase, the recombinants exhibited sPLA2 activity when analyzed by means of Langmuir dilauroylphosphatidylcholine monolayers as substrate. Both enzymes have a similar surface pressure-activity profile when compared with non-recombinant purified isoforms. To our knowledge, this is the first time that analysis of optimal lateral pressure of substrate monolayers by using the surface barostat technique is performed on recombinant sPLA2s.
Fil: Yunes Quartino, Pablo Javier. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Química Biológica; Argentina. 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: Barra, Jose Luis. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Química Biológica; Argentina. 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
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. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Química Biológica; Argentina
description Bothrops diporus is a very common viper in Argentina. At present, no complete sequence of secreted phospholipase A2 (sPLA2) from this snake has been reported. We have cloned two sPLA2 isoenzymes as well as a putative sPLA2-like myotoxin from venom gland. The two sPLA2 were expressed as inclusion bodies in Escherichia coli with an N-terminal tag of ubiquitin. After in vitro renaturation and cleavage step, using an ubiquitin specific peptidase, the recombinants exhibited sPLA2 activity when analyzed by means of Langmuir dilauroylphosphatidylcholine monolayers as substrate. Both enzymes have a similar surface pressure-activity profile when compared with non-recombinant purified isoforms. To our knowledge, this is the first time that analysis of optimal lateral pressure of substrate monolayers by using the surface barostat technique is performed on recombinant sPLA2s.
publishDate 2012
dc.date.none.fl_str_mv 2012-10
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/11336/198118
Yunes Quartino, Pablo Javier; Barra, Jose Luis; Fidelio, Gerardo Daniel; Cloning and functional expression of secreted phospholipases A2 from Bothrops diporus (Yarará Chica); Academic Press Inc Elsevier Science; Biochemical and Biophysical Research Communications; 427; 2; 10-2012; 321-325
0006-291X
CONICET Digital
CONICET
url http://hdl.handle.net/11336/198118
identifier_str_mv Yunes Quartino, Pablo Javier; Barra, Jose Luis; Fidelio, Gerardo Daniel; Cloning and functional expression of secreted phospholipases A2 from Bothrops diporus (Yarará Chica); Academic Press Inc Elsevier Science; Biochemical and Biophysical Research Communications; 427; 2; 10-2012; 321-325
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/http://www.sciencedirect.com/science/article/pii/S0006291X12017913
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bbrc.2012.09.051
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
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_ 1842981228307808256
score 12.48226

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