Purification and functional characterization of two fibrinogenolytic enzymes from Bothrops alternatus venom

Detalhes bibliográficos
Autor(a) principal: Costa,J. O.
Data de Publicação: 2007
Outros Autores: Petric,C. B., Hamaguchi,A., Homsi-Brandeburgo,M. I., Oliveira,C. Z., Soares,A. M., Oliveira,F.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: The Journal of venomous animals and toxins including tropical diseases (Online)
Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992007000300007
Resumo: Two fibrinogenolytic enzymes, Bothrops alternatus metalloprotease isoform (BaltMP)-I and II, were purified from Bothrops alternatus venom using Diethylaminoethyl (DEAE) Sephacel, Sephadex G-75 and Heparin-Agarose column chromatography. Purified BaltMP-I and II ran as single protein bands on analytical polyacrylamide gel electrophoresis and showed molecular weights of 29000 and 36000, respectively, under reducing conditions in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). BaltMP-II, but not BaltMP-I, displayed blood-clotting activity in bovine plasma, which was about 10-fold higher than that of the crude venom. Both enzymes were proteolytically active against bovine fibrinogen as substrate. When fibrinogen and each enzyme were incubated at 37°C, at a ratio of 1:100 (w/w), BaltMP-II cleaved preferentially the Aalpha -chain and more slowly the Bbeta -chain. The action of BaltMP-I was similar, but lower. None of the proteases degraded the gamma-chain of fibrinogen. The fibrinogenolytic activity of the enzymes was inhibited by 1,10-phenanthroline, suggesting they are metalloproteases. Since both enzymes were found to cause defibrinogenation when intraperitoneally (i.p.) administered to mice, they can be of medical interest as a therapeutic agent in the treatment and prevention of arterial thrombosis.
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spelling Purification and functional characterization of two fibrinogenolytic enzymes from Bothrops alternatus venomsnake venomBothrops alternatusmetalloproteasesfunctional characterizationfibrinogenolytic activitydefibrinogenation in vivoTwo fibrinogenolytic enzymes, Bothrops alternatus metalloprotease isoform (BaltMP)-I and II, were purified from Bothrops alternatus venom using Diethylaminoethyl (DEAE) Sephacel, Sephadex G-75 and Heparin-Agarose column chromatography. Purified BaltMP-I and II ran as single protein bands on analytical polyacrylamide gel electrophoresis and showed molecular weights of 29000 and 36000, respectively, under reducing conditions in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). BaltMP-II, but not BaltMP-I, displayed blood-clotting activity in bovine plasma, which was about 10-fold higher than that of the crude venom. Both enzymes were proteolytically active against bovine fibrinogen as substrate. When fibrinogen and each enzyme were incubated at 37°C, at a ratio of 1:100 (w/w), BaltMP-II cleaved preferentially the Aalpha -chain and more slowly the Bbeta -chain. The action of BaltMP-I was similar, but lower. None of the proteases degraded the gamma-chain of fibrinogen. The fibrinogenolytic activity of the enzymes was inhibited by 1,10-phenanthroline, suggesting they are metalloproteases. Since both enzymes were found to cause defibrinogenation when intraperitoneally (i.p.) administered to mice, they can be of medical interest as a therapeutic agent in the treatment and prevention of arterial thrombosis.Centro de Estudos de Venenos e Animais Peçonhentos (CEVAP/UNESP)2007-01-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992007000300007Journal of Venomous Animals and Toxins including Tropical Diseases v.13 n.3 2007reponame:The Journal of venomous animals and toxins including tropical diseases (Online)instname:Universidade Estadual Paulista (UNESP)instacron:UNESP10.1590/S1678-91992007000300007info:eu-repo/semantics/openAccessCosta,J. O.Petric,C. B.Hamaguchi,A.Homsi-Brandeburgo,M. I.Oliveira,C. Z.Soares,A. M.Oliveira,F.eng2007-09-17T00:00:00Zoai:scielo:S1678-91992007000300007Revistahttp://www.scielo.br/jvatitdPUBhttps://old.scielo.br/oai/scielo-oai.php||editorial@jvat.org.br1678-91991678-9180opendoar:2007-09-17T00:00The Journal of venomous animals and toxins including tropical diseases (Online) - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Purification and functional characterization of two fibrinogenolytic enzymes from Bothrops alternatus venom
title Purification and functional characterization of two fibrinogenolytic enzymes from Bothrops alternatus venom
spellingShingle Purification and functional characterization of two fibrinogenolytic enzymes from Bothrops alternatus venom
Costa,J. O.
snake venom
Bothrops alternatus
metalloproteases
functional characterization
fibrinogenolytic activity
defibrinogenation in vivo
title_short Purification and functional characterization of two fibrinogenolytic enzymes from Bothrops alternatus venom
title_full Purification and functional characterization of two fibrinogenolytic enzymes from Bothrops alternatus venom
title_fullStr Purification and functional characterization of two fibrinogenolytic enzymes from Bothrops alternatus venom
title_full_unstemmed Purification and functional characterization of two fibrinogenolytic enzymes from Bothrops alternatus venom
title_sort Purification and functional characterization of two fibrinogenolytic enzymes from Bothrops alternatus venom
author Costa,J. O.
author_facet Costa,J. O.
Petric,C. B.
Hamaguchi,A.
Homsi-Brandeburgo,M. I.
Oliveira,C. Z.
Soares,A. M.
Oliveira,F.
author_role author
author2 Petric,C. B.
Hamaguchi,A.
Homsi-Brandeburgo,M. I.
Oliveira,C. Z.
Soares,A. M.
Oliveira,F.
author2_role author
author
author
author
author
author
dc.contributor.author.fl_str_mv Costa,J. O.
Petric,C. B.
Hamaguchi,A.
Homsi-Brandeburgo,M. I.
Oliveira,C. Z.
Soares,A. M.
Oliveira,F.
dc.subject.por.fl_str_mv snake venom
Bothrops alternatus
metalloproteases
functional characterization
fibrinogenolytic activity
defibrinogenation in vivo
topic snake venom
Bothrops alternatus
metalloproteases
functional characterization
fibrinogenolytic activity
defibrinogenation in vivo
description Two fibrinogenolytic enzymes, Bothrops alternatus metalloprotease isoform (BaltMP)-I and II, were purified from Bothrops alternatus venom using Diethylaminoethyl (DEAE) Sephacel, Sephadex G-75 and Heparin-Agarose column chromatography. Purified BaltMP-I and II ran as single protein bands on analytical polyacrylamide gel electrophoresis and showed molecular weights of 29000 and 36000, respectively, under reducing conditions in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). BaltMP-II, but not BaltMP-I, displayed blood-clotting activity in bovine plasma, which was about 10-fold higher than that of the crude venom. Both enzymes were proteolytically active against bovine fibrinogen as substrate. When fibrinogen and each enzyme were incubated at 37°C, at a ratio of 1:100 (w/w), BaltMP-II cleaved preferentially the Aalpha -chain and more slowly the Bbeta -chain. The action of BaltMP-I was similar, but lower. None of the proteases degraded the gamma-chain of fibrinogen. The fibrinogenolytic activity of the enzymes was inhibited by 1,10-phenanthroline, suggesting they are metalloproteases. Since both enzymes were found to cause defibrinogenation when intraperitoneally (i.p.) administered to mice, they can be of medical interest as a therapeutic agent in the treatment and prevention of arterial thrombosis.
publishDate 2007
dc.date.none.fl_str_mv 2007-01-01
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992007000300007
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992007000300007
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 10.1590/S1678-91992007000300007
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv text/html
dc.publisher.none.fl_str_mv Centro de Estudos de Venenos e Animais Peçonhentos (CEVAP/UNESP)
publisher.none.fl_str_mv Centro de Estudos de Venenos e Animais Peçonhentos (CEVAP/UNESP)
dc.source.none.fl_str_mv Journal of Venomous Animals and Toxins including Tropical Diseases v.13 n.3 2007
reponame:The Journal of venomous animals and toxins including tropical diseases (Online)
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str The Journal of venomous animals and toxins including tropical diseases (Online)
collection The Journal of venomous animals and toxins including tropical diseases (Online)
repository.name.fl_str_mv The Journal of venomous animals and toxins including tropical diseases (Online) - Universidade Estadual Paulista (UNESP)
repository.mail.fl_str_mv ||editorial@jvat.org.br
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