A prothrombin activator from Bothrops erythromelas (jararaca-da-seca) snake venom: characterization and molecular cloning

Autores
Silva, Márcia B.; Schattner, Mirta Ana; Ramos, Celso R. R.; Junqueira de Azevedo, Inácio L. M.; Guarnieri, Míriam C.; Lazzari, María Ángela; Sampaio, Claudio A. M.; Pozner, Roberto Gabriel; Ventura, Janaina S.; Ho, Paulo L.; Chudzinski Tavassi, Ana M.
Año de publicación
2003
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
A novel prothrombin activator enzyme, which we have named 'berythractivase', was isolated from Bothrops erythromelas (jararaca-da-seca) snake venom. Berythractivase was purified by a single cation-exchange-chromatography step on a Resource S (Amersham Biosciences) column. The overall purification (31-fold) indicates that berythractivase comprises about 5% of the crude venom. It is a single-chain protein with a molecular mass of 78 kDa. SDS/PAGE of prothrombin after activation by berythractivase showed fragment patterns similar to those generated by group A prothrombin activators, which convert prothrombin into meizothrombin, independent of the prothrombinase complex. Chelating agents, such as EDTA and o-phenanthroline, rapidly inhibited the enzymic activity of berythractivase, like a typical metalloproteinase. Human fibrinogen Aα-chain was slowly digested only after longer incubation with berythractivase, and no effect on the β- or γ-chains was observed. Berythractivase was also capable of triggering endothelial proinfiammatory and procoagulant cell responses, von Willebrand factor was released, and the surface expression of both intracellular adhesion molecule-1 and E-selectin was up-regulated by berythractivase in cultured human umbilical-vein endothelial cells. The complete berythractivase cDNA was cloned from a B. erythromelas venom-gland cDNA library. The cDNA sequence possesses 2330 bp and encodes a preproprotein with significant sequence similarity to many other mature metalloproteinases reported from snake venoms. Berythractivase contains metalloproteinase, desintegrin-like and cysteine-rich domains. However, berythractivase did not elicit any haemorrhagic response. These results show that, although the primary structure of berythractivase is related to that of snake-venom haemorrhagic metalloproteinases and functionally similar to group A prothrombin activators, it is a prothrombin activator devoid of haemorrhagic activity. This is a feature not observed for most of the snake venom metalloproteinases, including the group A prothrombin activators.
Fil: Silva, Márcia B.. Universidade Federal de Pernambuco; Brasil. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
Fil: Schattner, Mirta Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; Argentina
Fil: Ramos, Celso R. R.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
Fil: Junqueira de Azevedo, Inácio L. M.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
Fil: Guarnieri, Míriam C.. Universidade Federal de Pernambuco; Brasil
Fil: Lazzari, María Ángela. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; Argentina
Fil: Sampaio, Claudio A. M.. Universidade Federal de Sao Paulo;
Fil: Pozner, Roberto Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; Argentina
Fil: Ventura, Janaina S.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
Fil: Ho, Paulo L.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
Fil: Chudzinski Tavassi, Ana M.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
Materia
Endothelium
Enzyme Activator
Fibrinogen
Cell Adhesion Molecules/Drug Effects
Von Willebrand Factor
Berythractivase
Coagulation
Thrombosis
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:11336/55306
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/55306
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling A prothrombin activator from Bothrops erythromelas (jararaca-da-seca) snake venom: characterization and molecular cloningSilva, Márcia B.Schattner, Mirta AnaRamos, Celso R. R.Junqueira de Azevedo, Inácio L. M.Guarnieri, Míriam C.Lazzari, María ÁngelaSampaio, Claudio A. M.Pozner, Roberto GabrielVentura, Janaina S.Ho, Paulo L.Chudzinski Tavassi, Ana M.EndotheliumEnzyme ActivatorFibrinogenCell Adhesion Molecules/Drug EffectsVon Willebrand FactorBerythractivaseCoagulationThrombosishttps://purl.org/becyt/ford/3.2https://purl.org/becyt/ford/3A novel prothrombin activator enzyme, which we have named 'berythractivase', was isolated from Bothrops erythromelas (jararaca-da-seca) snake venom. Berythractivase was purified by a single cation-exchange-chromatography step on a Resource S (Amersham Biosciences) column. The overall purification (31-fold) indicates that berythractivase comprises about 5% of the crude venom. It is a single-chain protein with a molecular mass of 78 kDa. SDS/PAGE of prothrombin after activation by berythractivase showed fragment patterns similar to those generated by group A prothrombin activators, which convert prothrombin into meizothrombin, independent of the prothrombinase complex. Chelating agents, such as EDTA and o-phenanthroline, rapidly inhibited the enzymic activity of berythractivase, like a typical metalloproteinase. Human fibrinogen Aα-chain was slowly digested only after longer incubation with berythractivase, and no effect on the β- or γ-chains was observed. Berythractivase was also capable of triggering endothelial proinfiammatory and procoagulant cell responses, von Willebrand factor was released, and the surface expression of both intracellular adhesion molecule-1 and E-selectin was up-regulated by berythractivase in cultured human umbilical-vein endothelial cells. The complete berythractivase cDNA was cloned from a B. erythromelas venom-gland cDNA library. The cDNA sequence possesses 2330 bp and encodes a preproprotein with significant sequence similarity to many other mature metalloproteinases reported from snake venoms. Berythractivase contains metalloproteinase, desintegrin-like and cysteine-rich domains. However, berythractivase did not elicit any haemorrhagic response. These results show that, although the primary structure of berythractivase is related to that of snake-venom haemorrhagic metalloproteinases and functionally similar to group A prothrombin activators, it is a prothrombin activator devoid of haemorrhagic activity. This is a feature not observed for most of the snake venom metalloproteinases, including the group A prothrombin activators.Fil: Silva, Márcia B.. Universidade Federal de Pernambuco; Brasil. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; BrasilFil: Schattner, Mirta Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; ArgentinaFil: Ramos, Celso R. R.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; BrasilFil: Junqueira de Azevedo, Inácio L. M.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; BrasilFil: Guarnieri, Míriam C.. Universidade Federal de Pernambuco; BrasilFil: Lazzari, María Ángela. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; ArgentinaFil: Sampaio, Claudio A. M.. Universidade Federal de Sao Paulo;Fil: Pozner, Roberto Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; ArgentinaFil: Ventura, Janaina S.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; BrasilFil: Ho, Paulo L.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; BrasilFil: Chudzinski Tavassi, Ana M.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; BrasilPortland Press2003-01info: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/11336/55306Silva, Márcia B.; Schattner, Mirta Ana; Ramos, Celso R. R.; Junqueira de Azevedo, Inácio L. M.; Guarnieri, Míriam C.; et al.; A prothrombin activator from Bothrops erythromelas (jararaca-da-seca) snake venom: characterization and molecular cloning; Portland Press; Biochemical Journal; 369; 1-2003; 129-1390264-60211470-8728CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1042/BJ20020449info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2026-01-14T12:57:25Zoai:ri.conicet.gov.ar:11336/55306instacron: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:34982026-01-14 12:57:25.454CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv A prothrombin activator from Bothrops erythromelas (jararaca-da-seca) snake venom: characterization and molecular cloning
title A prothrombin activator from Bothrops erythromelas (jararaca-da-seca) snake venom: characterization and molecular cloning
spellingShingle A prothrombin activator from Bothrops erythromelas (jararaca-da-seca) snake venom: characterization and molecular cloning
Silva, Márcia B.
Endothelium
Enzyme Activator
Fibrinogen
Cell Adhesion Molecules/Drug Effects
Von Willebrand Factor
Berythractivase
Coagulation
Thrombosis
title_short A prothrombin activator from Bothrops erythromelas (jararaca-da-seca) snake venom: characterization and molecular cloning
title_full A prothrombin activator from Bothrops erythromelas (jararaca-da-seca) snake venom: characterization and molecular cloning
title_fullStr A prothrombin activator from Bothrops erythromelas (jararaca-da-seca) snake venom: characterization and molecular cloning
title_full_unstemmed A prothrombin activator from Bothrops erythromelas (jararaca-da-seca) snake venom: characterization and molecular cloning
title_sort A prothrombin activator from Bothrops erythromelas (jararaca-da-seca) snake venom: characterization and molecular cloning
dc.creator.none.fl_str_mv Silva, Márcia B.
Schattner, Mirta Ana
Ramos, Celso R. R.
Junqueira de Azevedo, Inácio L. M.
Guarnieri, Míriam C.
Lazzari, María Ángela
Sampaio, Claudio A. M.
Pozner, Roberto Gabriel
Ventura, Janaina S.
Ho, Paulo L.
Chudzinski Tavassi, Ana M.
author Silva, Márcia B.
author_facet Silva, Márcia B.
Schattner, Mirta Ana
Ramos, Celso R. R.
Junqueira de Azevedo, Inácio L. M.
Guarnieri, Míriam C.
Lazzari, María Ángela
Sampaio, Claudio A. M.
Pozner, Roberto Gabriel
Ventura, Janaina S.
Ho, Paulo L.
Chudzinski Tavassi, Ana M.
author_role author
author2 Schattner, Mirta Ana
Ramos, Celso R. R.
Junqueira de Azevedo, Inácio L. M.
Guarnieri, Míriam C.
Lazzari, María Ángela
Sampaio, Claudio A. M.
Pozner, Roberto Gabriel
Ventura, Janaina S.
Ho, Paulo L.
Chudzinski Tavassi, Ana M.
author2_role author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Endothelium
Enzyme Activator
Fibrinogen
Cell Adhesion Molecules/Drug Effects
Von Willebrand Factor
Berythractivase
Coagulation
Thrombosis
topic Endothelium
Enzyme Activator
Fibrinogen
Cell Adhesion Molecules/Drug Effects
Von Willebrand Factor
Berythractivase
Coagulation
Thrombosis
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.2
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv A novel prothrombin activator enzyme, which we have named 'berythractivase', was isolated from Bothrops erythromelas (jararaca-da-seca) snake venom. Berythractivase was purified by a single cation-exchange-chromatography step on a Resource S (Amersham Biosciences) column. The overall purification (31-fold) indicates that berythractivase comprises about 5% of the crude venom. It is a single-chain protein with a molecular mass of 78 kDa. SDS/PAGE of prothrombin after activation by berythractivase showed fragment patterns similar to those generated by group A prothrombin activators, which convert prothrombin into meizothrombin, independent of the prothrombinase complex. Chelating agents, such as EDTA and o-phenanthroline, rapidly inhibited the enzymic activity of berythractivase, like a typical metalloproteinase. Human fibrinogen Aα-chain was slowly digested only after longer incubation with berythractivase, and no effect on the β- or γ-chains was observed. Berythractivase was also capable of triggering endothelial proinfiammatory and procoagulant cell responses, von Willebrand factor was released, and the surface expression of both intracellular adhesion molecule-1 and E-selectin was up-regulated by berythractivase in cultured human umbilical-vein endothelial cells. The complete berythractivase cDNA was cloned from a B. erythromelas venom-gland cDNA library. The cDNA sequence possesses 2330 bp and encodes a preproprotein with significant sequence similarity to many other mature metalloproteinases reported from snake venoms. Berythractivase contains metalloproteinase, desintegrin-like and cysteine-rich domains. However, berythractivase did not elicit any haemorrhagic response. These results show that, although the primary structure of berythractivase is related to that of snake-venom haemorrhagic metalloproteinases and functionally similar to group A prothrombin activators, it is a prothrombin activator devoid of haemorrhagic activity. This is a feature not observed for most of the snake venom metalloproteinases, including the group A prothrombin activators.
Fil: Silva, Márcia B.. Universidade Federal de Pernambuco; Brasil. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
Fil: Schattner, Mirta Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; Argentina
Fil: Ramos, Celso R. R.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
Fil: Junqueira de Azevedo, Inácio L. M.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
Fil: Guarnieri, Míriam C.. Universidade Federal de Pernambuco; Brasil
Fil: Lazzari, María Ángela. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; Argentina
Fil: Sampaio, Claudio A. M.. Universidade Federal de Sao Paulo;
Fil: Pozner, Roberto Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; Argentina
Fil: Ventura, Janaina S.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
Fil: Ho, Paulo L.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
Fil: Chudzinski Tavassi, Ana M.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
description A novel prothrombin activator enzyme, which we have named 'berythractivase', was isolated from Bothrops erythromelas (jararaca-da-seca) snake venom. Berythractivase was purified by a single cation-exchange-chromatography step on a Resource S (Amersham Biosciences) column. The overall purification (31-fold) indicates that berythractivase comprises about 5% of the crude venom. It is a single-chain protein with a molecular mass of 78 kDa. SDS/PAGE of prothrombin after activation by berythractivase showed fragment patterns similar to those generated by group A prothrombin activators, which convert prothrombin into meizothrombin, independent of the prothrombinase complex. Chelating agents, such as EDTA and o-phenanthroline, rapidly inhibited the enzymic activity of berythractivase, like a typical metalloproteinase. Human fibrinogen Aα-chain was slowly digested only after longer incubation with berythractivase, and no effect on the β- or γ-chains was observed. Berythractivase was also capable of triggering endothelial proinfiammatory and procoagulant cell responses, von Willebrand factor was released, and the surface expression of both intracellular adhesion molecule-1 and E-selectin was up-regulated by berythractivase in cultured human umbilical-vein endothelial cells. The complete berythractivase cDNA was cloned from a B. erythromelas venom-gland cDNA library. The cDNA sequence possesses 2330 bp and encodes a preproprotein with significant sequence similarity to many other mature metalloproteinases reported from snake venoms. Berythractivase contains metalloproteinase, desintegrin-like and cysteine-rich domains. However, berythractivase did not elicit any haemorrhagic response. These results show that, although the primary structure of berythractivase is related to that of snake-venom haemorrhagic metalloproteinases and functionally similar to group A prothrombin activators, it is a prothrombin activator devoid of haemorrhagic activity. This is a feature not observed for most of the snake venom metalloproteinases, including the group A prothrombin activators.
publishDate 2003
dc.date.none.fl_str_mv 2003-01
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/11336/55306
Silva, Márcia B.; Schattner, Mirta Ana; Ramos, Celso R. R.; Junqueira de Azevedo, Inácio L. M.; Guarnieri, Míriam C.; et al.; A prothrombin activator from Bothrops erythromelas (jararaca-da-seca) snake venom: characterization and molecular cloning; Portland Press; Biochemical Journal; 369; 1-2003; 129-139
0264-6021
1470-8728
CONICET Digital
CONICET
url http://hdl.handle.net/11336/55306
identifier_str_mv Silva, Márcia B.; Schattner, Mirta Ana; Ramos, Celso R. R.; Junqueira de Azevedo, Inácio L. M.; Guarnieri, Míriam C.; et al.; A prothrombin activator from Bothrops erythromelas (jararaca-da-seca) snake venom: characterization and molecular cloning; Portland Press; Biochemical Journal; 369; 1-2003; 129-139
0264-6021
1470-8728
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1042/BJ20020449
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 Portland Press
publisher.none.fl_str_mv Portland Press
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 13.071171

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