Characterization of phenoloxidase activity from spider Polybetes pythagoricus hemocyanin

Autores
Laino, Aldana; Lavarias, Sabrina Maria Luisa; Suarez, Gustavo; Lino, Agustina Berenice; Cunningham, Monica Liliana
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Hemocyanin of the spider Polybetes pythagoricus, in addition to its typical role as an oxygen transporter, also exhibits a phenoloxidase activity induced by micellar concentrations of SDS. In the present work, we found the kinetic parameters Km and Vmax of Polybetes pythagoricus hemocyanin (PpHc) PO activity to be 0.407 mM and 0.081 µmolmin−1mg protein−1, respectively. Dopamine was used as the substrate with SDS at a final concentration of 10 mM and a 30‐min incubation at 25°C. Conformational changes in Hc associated with the SDS treatment were analyzed using far‐UV circular dichroism, intrinsic fluorescence and absorption spectroscopy. The secondary and tertiary structural changes of PpHc induced by SDS led to increases in α‐helical content and tryptophan fluorescence intensity. A reduction in the absorption spectrum at 340 nm in the presence of SDS was also observed. These results suggest that the SDS‐induced PO activity of PpHc can be ascribed to conformational changes in the local environment of the typer‐3 copper active site.
Fil: Laino, Aldana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; Argentina
Fil: Lavarias, Sabrina Maria Luisa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Limnología "Dr. Raúl A. Ringuelet". Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Limnología; Argentina
Fil: Suarez, Gustavo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; Argentina
Fil: Lino, Agustina Berenice. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Laboratorio de Investigación en Osteospatías y Metabolismo Mineral; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Cunningham, Monica Liliana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; Argentina
Materia
Phenoloxidase
Spider
Hemocyanin
Activity
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:11336/48854
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/48854
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Characterization of phenoloxidase activity from spider Polybetes pythagoricus hemocyaninLaino, AldanaLavarias, Sabrina Maria LuisaSuarez, GustavoLino, Agustina BereniceCunningham, Monica LilianaPhenoloxidaseSpiderHemocyaninActivityhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Hemocyanin of the spider Polybetes pythagoricus, in addition to its typical role as an oxygen transporter, also exhibits a phenoloxidase activity induced by micellar concentrations of SDS. In the present work, we found the kinetic parameters Km and Vmax of Polybetes pythagoricus hemocyanin (PpHc) PO activity to be 0.407 mM and 0.081 µmolmin−1mg protein−1, respectively. Dopamine was used as the substrate with SDS at a final concentration of 10 mM and a 30‐min incubation at 25°C. Conformational changes in Hc associated with the SDS treatment were analyzed using far‐UV circular dichroism, intrinsic fluorescence and absorption spectroscopy. The secondary and tertiary structural changes of PpHc induced by SDS led to increases in α‐helical content and tryptophan fluorescence intensity. A reduction in the absorption spectrum at 340 nm in the presence of SDS was also observed. These results suggest that the SDS‐induced PO activity of PpHc can be ascribed to conformational changes in the local environment of the typer‐3 copper active site.Fil: Laino, Aldana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; ArgentinaFil: Lavarias, Sabrina Maria Luisa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Limnología "Dr. Raúl A. Ringuelet". Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Limnología; ArgentinaFil: Suarez, Gustavo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; ArgentinaFil: Lino, Agustina Berenice. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Laboratorio de Investigación en Osteospatías y Metabolismo Mineral; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Cunningham, Monica Liliana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; ArgentinaWiley-liss, Div John Wiley & Sons Inc2015-10info: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/11336/48854Laino, Aldana; Lavarias, Sabrina Maria Luisa; Suarez, Gustavo; Lino, Agustina Berenice; Cunningham, Monica Liliana; Characterization of phenoloxidase activity from spider Polybetes pythagoricus hemocyanin; Wiley-liss, Div John Wiley & Sons Inc; Journal of Experimental Zoology Part A: Ecological Genetics and Physiology; 323; 8; 10-2015; 547-5551932-5223CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1002/jez.1947info:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/abs/10.1002/jez.1947info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T15:44:05Zoai:ri.conicet.gov.ar:11336/48854instacron: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:34982025-10-15 15:44:05.607CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Characterization of phenoloxidase activity from spider Polybetes pythagoricus hemocyanin
title Characterization of phenoloxidase activity from spider Polybetes pythagoricus hemocyanin
spellingShingle Characterization of phenoloxidase activity from spider Polybetes pythagoricus hemocyanin
Laino, Aldana
Phenoloxidase
Spider
Hemocyanin
Activity
title_short Characterization of phenoloxidase activity from spider Polybetes pythagoricus hemocyanin
title_full Characterization of phenoloxidase activity from spider Polybetes pythagoricus hemocyanin
title_fullStr Characterization of phenoloxidase activity from spider Polybetes pythagoricus hemocyanin
title_full_unstemmed Characterization of phenoloxidase activity from spider Polybetes pythagoricus hemocyanin
title_sort Characterization of phenoloxidase activity from spider Polybetes pythagoricus hemocyanin
dc.creator.none.fl_str_mv Laino, Aldana
Lavarias, Sabrina Maria Luisa
Suarez, Gustavo
Lino, Agustina Berenice
Cunningham, Monica Liliana
author Laino, Aldana
author_facet Laino, Aldana
Lavarias, Sabrina Maria Luisa
Suarez, Gustavo
Lino, Agustina Berenice
Cunningham, Monica Liliana
author_role author
author2 Lavarias, Sabrina Maria Luisa
Suarez, Gustavo
Lino, Agustina Berenice
Cunningham, Monica Liliana
author2_role author
author
author
author
dc.subject.none.fl_str_mv Phenoloxidase
Spider
Hemocyanin
Activity
topic Phenoloxidase
Spider
Hemocyanin
Activity
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Hemocyanin of the spider Polybetes pythagoricus, in addition to its typical role as an oxygen transporter, also exhibits a phenoloxidase activity induced by micellar concentrations of SDS. In the present work, we found the kinetic parameters Km and Vmax of Polybetes pythagoricus hemocyanin (PpHc) PO activity to be 0.407 mM and 0.081 µmolmin−1mg protein−1, respectively. Dopamine was used as the substrate with SDS at a final concentration of 10 mM and a 30‐min incubation at 25°C. Conformational changes in Hc associated with the SDS treatment were analyzed using far‐UV circular dichroism, intrinsic fluorescence and absorption spectroscopy. The secondary and tertiary structural changes of PpHc induced by SDS led to increases in α‐helical content and tryptophan fluorescence intensity. A reduction in the absorption spectrum at 340 nm in the presence of SDS was also observed. These results suggest that the SDS‐induced PO activity of PpHc can be ascribed to conformational changes in the local environment of the typer‐3 copper active site.
Fil: Laino, Aldana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; Argentina
Fil: Lavarias, Sabrina Maria Luisa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Limnología "Dr. Raúl A. Ringuelet". Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Limnología; Argentina
Fil: Suarez, Gustavo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; Argentina
Fil: Lino, Agustina Berenice. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Laboratorio de Investigación en Osteospatías y Metabolismo Mineral; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Cunningham, Monica Liliana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; Argentina
description Hemocyanin of the spider Polybetes pythagoricus, in addition to its typical role as an oxygen transporter, also exhibits a phenoloxidase activity induced by micellar concentrations of SDS. In the present work, we found the kinetic parameters Km and Vmax of Polybetes pythagoricus hemocyanin (PpHc) PO activity to be 0.407 mM and 0.081 µmolmin−1mg protein−1, respectively. Dopamine was used as the substrate with SDS at a final concentration of 10 mM and a 30‐min incubation at 25°C. Conformational changes in Hc associated with the SDS treatment were analyzed using far‐UV circular dichroism, intrinsic fluorescence and absorption spectroscopy. The secondary and tertiary structural changes of PpHc induced by SDS led to increases in α‐helical content and tryptophan fluorescence intensity. A reduction in the absorption spectrum at 340 nm in the presence of SDS was also observed. These results suggest that the SDS‐induced PO activity of PpHc can be ascribed to conformational changes in the local environment of the typer‐3 copper active site.
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/11336/48854
Laino, Aldana; Lavarias, Sabrina Maria Luisa; Suarez, Gustavo; Lino, Agustina Berenice; Cunningham, Monica Liliana; Characterization of phenoloxidase activity from spider Polybetes pythagoricus hemocyanin; Wiley-liss, Div John Wiley & Sons Inc; Journal of Experimental Zoology Part A: Ecological Genetics and Physiology; 323; 8; 10-2015; 547-555
1932-5223
CONICET Digital
CONICET
url http://hdl.handle.net/11336/48854
identifier_str_mv Laino, Aldana; Lavarias, Sabrina Maria Luisa; Suarez, Gustavo; Lino, Agustina Berenice; Cunningham, Monica Liliana; Characterization of phenoloxidase activity from spider Polybetes pythagoricus hemocyanin; Wiley-liss, Div John Wiley & Sons Inc; Journal of Experimental Zoology Part A: Ecological Genetics and Physiology; 323; 8; 10-2015; 547-555
1932-5223
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1002/jez.1947
info:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/abs/10.1002/jez.1947
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
dc.publisher.none.fl_str_mv Wiley-liss, Div John Wiley & Sons Inc
publisher.none.fl_str_mv Wiley-liss, Div John Wiley & Sons Inc
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_ 1846083544522162176
score 13.22299

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