Purification and functional characterization of two fibrinogenolytic enzymes from Bothrops alternatus venom
Autor(a) principal: | |
---|---|
Data de Publicação: | 2007 |
Outros Autores: | , , , , , |
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. |
id |
UNESP-11_cd44a23614ae1b59242419f036d3ae35 |
---|---|
oai_identifier_str |
oai:scielo:S1678-91992007000300007 |
network_acronym_str |
UNESP-11 |
network_name_str |
The Journal of venomous animals and toxins including tropical diseases (Online) |
repository_id_str |
|
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 |
_version_ |
1748958537971662848 |