Proteolytic, edematogenic and myotoxic activities of a hemorrhagic metalloproteinase isolated from Bothrops alternatus venom

Autores
Gay, Claudia Carolina; Leiva, Laura Cristina Ana; Maruñak, Silvana; Teibler, Gladys Pamela; Acosta de Pérez, Ofelia
Año de publicación
2005
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
A hemorrhagic metalloproteinase has been isolated from Bothrops alternatus venom from specimens that inhabit the north-east region of Argentina. The present study aimed at evaluating the proteolytic, hemorrhagic, edematogenic and myotoxic activities of the purified metalloproteinase, in order to consider its participation on the phatophysiology of the intoxication by Bothrops alternatus venom. The hemorrhagic metalloproteinase was isolated by a combination of DEAE-Cellulose chromatography and gel filtration on Sephadex G-75. The enzyme showed a molecular mass around 55 kDa, it exhibited a hemorrhagic activity with a minimal hemorrhagic dose of 1.9 μg, almost two fold minor than the whole venom (3.6 μg). The enzyme showed a weak proteolytic activity on casein (18.72 U/mg enzyme), similar to the one exhibited by the whole venom (20 U/mg venom). Besides, the ability to degrade casein could be detected by SDS-PAGE; β-casein was the fraction that showed the higher degradation, followed by αs1-casein and κ-casein degradation. The hemorrhagic metalloproteinase rapidly hydrolysed the Aα-chain of fibrinogen, followed by Bβ-chain degradation and leaving the γ-chain unaffected. Proteolytic activities were inhibited by EDTA whereas they were not inhibited by benzamidine and PMSF. The metalloproteinase showed several polypeptides chains after autocatalytic processing, including a chain of 28 kDa, it could be the processed disintegrin-like and cysteine-rich domains. The isolated enzyme exhibited myotoxic activity with high CK levels at 6 h, due to local ischemia resulting of its hemorrhagic activity, and a significant edema-inducing effect (MED=1.3 μg), corroborated both results by the histological observations of samples of gastrocnemius muscle. These findings showed that this hemorrhagic metalloproteinase, possesses high edematogenic and myotoxic activities and, in despite of exhibiting a weak proteolytic activity, it is able to degrade fibrinogen. So, this enzyme would contribute markedly to the phatophysiology of the bothropic envenomation.
Fil: Gay, Claudia Carolina. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas y Naturales y Agrimensura; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Leiva, Laura Cristina Ana. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas y Naturales y Agrimensura; Argentina
Fil: Maruñak, Silvana. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; Argentina
Fil: Teibler, Gladys Pamela. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; Argentina
Fil: Acosta de Pérez, Ofelia. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; Argentina
Materia
Hemorrhagin
Metalloproteinase
Snake Venom
Bothrops Alternatus
Nivel de accesibilidad
acceso abierto
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:11336/44666
Acceder
id CONICETDig_a93914a8f33218e9295c519042b69060
oai_identifier_str oai:ri.conicet.gov.ar:11336/44666
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Proteolytic, edematogenic and myotoxic activities of a hemorrhagic metalloproteinase isolated from Bothrops alternatus venomGay, Claudia CarolinaLeiva, Laura Cristina AnaMaruñak, SilvanaTeibler, Gladys PamelaAcosta de Pérez, OfeliaHemorrhaginMetalloproteinaseSnake VenomBothrops Alternatushttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1A hemorrhagic metalloproteinase has been isolated from Bothrops alternatus venom from specimens that inhabit the north-east region of Argentina. The present study aimed at evaluating the proteolytic, hemorrhagic, edematogenic and myotoxic activities of the purified metalloproteinase, in order to consider its participation on the phatophysiology of the intoxication by Bothrops alternatus venom. The hemorrhagic metalloproteinase was isolated by a combination of DEAE-Cellulose chromatography and gel filtration on Sephadex G-75. The enzyme showed a molecular mass around 55 kDa, it exhibited a hemorrhagic activity with a minimal hemorrhagic dose of 1.9 μg, almost two fold minor than the whole venom (3.6 μg). The enzyme showed a weak proteolytic activity on casein (18.72 U/mg enzyme), similar to the one exhibited by the whole venom (20 U/mg venom). Besides, the ability to degrade casein could be detected by SDS-PAGE; β-casein was the fraction that showed the higher degradation, followed by αs1-casein and κ-casein degradation. The hemorrhagic metalloproteinase rapidly hydrolysed the Aα-chain of fibrinogen, followed by Bβ-chain degradation and leaving the γ-chain unaffected. Proteolytic activities were inhibited by EDTA whereas they were not inhibited by benzamidine and PMSF. The metalloproteinase showed several polypeptides chains after autocatalytic processing, including a chain of 28 kDa, it could be the processed disintegrin-like and cysteine-rich domains. The isolated enzyme exhibited myotoxic activity with high CK levels at 6 h, due to local ischemia resulting of its hemorrhagic activity, and a significant edema-inducing effect (MED=1.3 μg), corroborated both results by the histological observations of samples of gastrocnemius muscle. These findings showed that this hemorrhagic metalloproteinase, possesses high edematogenic and myotoxic activities and, in despite of exhibiting a weak proteolytic activity, it is able to degrade fibrinogen. So, this enzyme would contribute markedly to the phatophysiology of the bothropic envenomation.Fil: Gay, Claudia Carolina. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas y Naturales y Agrimensura; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Leiva, Laura Cristina Ana. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas y Naturales y Agrimensura; ArgentinaFil: Maruñak, Silvana. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; ArgentinaFil: Teibler, Gladys Pamela. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; ArgentinaFil: Acosta de Pérez, Ofelia. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; ArgentinaPergamon-Elsevier Science Ltd2005-10info: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/11336/44666Gay, Claudia Carolina; Leiva, Laura Cristina Ana; Maruñak, Silvana; Teibler, Gladys Pamela; Acosta de Pérez, Ofelia; Proteolytic, edematogenic and myotoxic activities of a hemorrhagic metalloproteinase isolated from Bothrops alternatus venom; Pergamon-Elsevier Science Ltd; Toxicon; 46; 5; 10-2005; 546-5540041-0101CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.toxicon.2005.06.019info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0041010105002308info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:41:53Zoai:ri.conicet.gov.ar:11336/44666instacron: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:34982025-09-29 10:41:53.875CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Proteolytic, edematogenic and myotoxic activities of a hemorrhagic metalloproteinase isolated from Bothrops alternatus venom
title Proteolytic, edematogenic and myotoxic activities of a hemorrhagic metalloproteinase isolated from Bothrops alternatus venom
spellingShingle Proteolytic, edematogenic and myotoxic activities of a hemorrhagic metalloproteinase isolated from Bothrops alternatus venom
Gay, Claudia Carolina
Hemorrhagin
Metalloproteinase
Snake Venom
Bothrops Alternatus
title_short Proteolytic, edematogenic and myotoxic activities of a hemorrhagic metalloproteinase isolated from Bothrops alternatus venom
title_full Proteolytic, edematogenic and myotoxic activities of a hemorrhagic metalloproteinase isolated from Bothrops alternatus venom
title_fullStr Proteolytic, edematogenic and myotoxic activities of a hemorrhagic metalloproteinase isolated from Bothrops alternatus venom
title_full_unstemmed Proteolytic, edematogenic and myotoxic activities of a hemorrhagic metalloproteinase isolated from Bothrops alternatus venom
title_sort Proteolytic, edematogenic and myotoxic activities of a hemorrhagic metalloproteinase isolated from Bothrops alternatus venom
dc.creator.none.fl_str_mv Gay, Claudia Carolina
Leiva, Laura Cristina Ana
Maruñak, Silvana
Teibler, Gladys Pamela
Acosta de Pérez, Ofelia
author Gay, Claudia Carolina
author_facet Gay, Claudia Carolina
Leiva, Laura Cristina Ana
Maruñak, Silvana
Teibler, Gladys Pamela
Acosta de Pérez, Ofelia
author_role author
author2 Leiva, Laura Cristina Ana
Maruñak, Silvana
Teibler, Gladys Pamela
Acosta de Pérez, Ofelia
author2_role author
author
author
author
dc.subject.none.fl_str_mv Hemorrhagin
Metalloproteinase
Snake Venom
Bothrops Alternatus
topic Hemorrhagin
Metalloproteinase
Snake Venom
Bothrops Alternatus
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv A hemorrhagic metalloproteinase has been isolated from Bothrops alternatus venom from specimens that inhabit the north-east region of Argentina. The present study aimed at evaluating the proteolytic, hemorrhagic, edematogenic and myotoxic activities of the purified metalloproteinase, in order to consider its participation on the phatophysiology of the intoxication by Bothrops alternatus venom. The hemorrhagic metalloproteinase was isolated by a combination of DEAE-Cellulose chromatography and gel filtration on Sephadex G-75. The enzyme showed a molecular mass around 55 kDa, it exhibited a hemorrhagic activity with a minimal hemorrhagic dose of 1.9 μg, almost two fold minor than the whole venom (3.6 μg). The enzyme showed a weak proteolytic activity on casein (18.72 U/mg enzyme), similar to the one exhibited by the whole venom (20 U/mg venom). Besides, the ability to degrade casein could be detected by SDS-PAGE; β-casein was the fraction that showed the higher degradation, followed by αs1-casein and κ-casein degradation. The hemorrhagic metalloproteinase rapidly hydrolysed the Aα-chain of fibrinogen, followed by Bβ-chain degradation and leaving the γ-chain unaffected. Proteolytic activities were inhibited by EDTA whereas they were not inhibited by benzamidine and PMSF. The metalloproteinase showed several polypeptides chains after autocatalytic processing, including a chain of 28 kDa, it could be the processed disintegrin-like and cysteine-rich domains. The isolated enzyme exhibited myotoxic activity with high CK levels at 6 h, due to local ischemia resulting of its hemorrhagic activity, and a significant edema-inducing effect (MED=1.3 μg), corroborated both results by the histological observations of samples of gastrocnemius muscle. These findings showed that this hemorrhagic metalloproteinase, possesses high edematogenic and myotoxic activities and, in despite of exhibiting a weak proteolytic activity, it is able to degrade fibrinogen. So, this enzyme would contribute markedly to the phatophysiology of the bothropic envenomation.
Fil: Gay, Claudia Carolina. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas y Naturales y Agrimensura; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Leiva, Laura Cristina Ana. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas y Naturales y Agrimensura; Argentina
Fil: Maruñak, Silvana. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; Argentina
Fil: Teibler, Gladys Pamela. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; Argentina
Fil: Acosta de Pérez, Ofelia. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; Argentina
description A hemorrhagic metalloproteinase has been isolated from Bothrops alternatus venom from specimens that inhabit the north-east region of Argentina. The present study aimed at evaluating the proteolytic, hemorrhagic, edematogenic and myotoxic activities of the purified metalloproteinase, in order to consider its participation on the phatophysiology of the intoxication by Bothrops alternatus venom. The hemorrhagic metalloproteinase was isolated by a combination of DEAE-Cellulose chromatography and gel filtration on Sephadex G-75. The enzyme showed a molecular mass around 55 kDa, it exhibited a hemorrhagic activity with a minimal hemorrhagic dose of 1.9 μg, almost two fold minor than the whole venom (3.6 μg). The enzyme showed a weak proteolytic activity on casein (18.72 U/mg enzyme), similar to the one exhibited by the whole venom (20 U/mg venom). Besides, the ability to degrade casein could be detected by SDS-PAGE; β-casein was the fraction that showed the higher degradation, followed by αs1-casein and κ-casein degradation. The hemorrhagic metalloproteinase rapidly hydrolysed the Aα-chain of fibrinogen, followed by Bβ-chain degradation and leaving the γ-chain unaffected. Proteolytic activities were inhibited by EDTA whereas they were not inhibited by benzamidine and PMSF. The metalloproteinase showed several polypeptides chains after autocatalytic processing, including a chain of 28 kDa, it could be the processed disintegrin-like and cysteine-rich domains. The isolated enzyme exhibited myotoxic activity with high CK levels at 6 h, due to local ischemia resulting of its hemorrhagic activity, and a significant edema-inducing effect (MED=1.3 μg), corroborated both results by the histological observations of samples of gastrocnemius muscle. These findings showed that this hemorrhagic metalloproteinase, possesses high edematogenic and myotoxic activities and, in despite of exhibiting a weak proteolytic activity, it is able to degrade fibrinogen. So, this enzyme would contribute markedly to the phatophysiology of the bothropic envenomation.
publishDate 2005
dc.date.none.fl_str_mv 2005-10
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/11336/44666
Gay, Claudia Carolina; Leiva, Laura Cristina Ana; Maruñak, Silvana; Teibler, Gladys Pamela; Acosta de Pérez, Ofelia; Proteolytic, edematogenic and myotoxic activities of a hemorrhagic metalloproteinase isolated from Bothrops alternatus venom; Pergamon-Elsevier Science Ltd; Toxicon; 46; 5; 10-2005; 546-554
0041-0101
CONICET Digital
CONICET
url http://hdl.handle.net/11336/44666
identifier_str_mv Gay, Claudia Carolina; Leiva, Laura Cristina Ana; Maruñak, Silvana; Teibler, Gladys Pamela; Acosta de Pérez, Ofelia; Proteolytic, edematogenic and myotoxic activities of a hemorrhagic metalloproteinase isolated from Bothrops alternatus venom; Pergamon-Elsevier Science Ltd; Toxicon; 46; 5; 10-2005; 546-554
0041-0101
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.toxicon.2005.06.019
info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0041010105002308
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
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
dc.publisher.none.fl_str_mv Pergamon-Elsevier Science Ltd
publisher.none.fl_str_mv Pergamon-Elsevier Science Ltd
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_ 1844614450811764736
score 13.070432

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