Purification and functional characterization of two fibrinogenolytic enzymes from Bothrops alternatus venom
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 analytic...
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| Format: | Article |
| Language: | English |
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SciELO
2007-01-01
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| Series: | Journal of Venomous Animals and Toxins including Tropical Diseases |
| Subjects: | |
| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992007000300007 |
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| author | J. O. Costa C. B. Petric A. Hamaguchi M. I. Homsi-Brandeburgo C. Z. Oliveira A. M. Soares F. Oliveira |
| author_facet | J. O. Costa C. B. Petric A. Hamaguchi M. I. Homsi-Brandeburgo C. Z. Oliveira A. M. Soares F. Oliveira |
| author_sort | J. O. Costa |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-04-12T07:03:52Z |
| format | Article |
| id | doaj.art-6fb72fe992b64298915b9eb097080460 |
| institution | Directory Open Access Journal |
| issn | 1678-9199 |
| language | English |
| last_indexed | 2024-04-12T07:03:52Z |
| publishDate | 2007-01-01 |
| publisher | SciELO |
| record_format | Article |
| series | Journal of Venomous Animals and Toxins including Tropical Diseases |
| spelling | doaj.art-6fb72fe992b64298915b9eb0970804602022-12-22T03:42:54ZengSciELOJournal of Venomous Animals and Toxins including Tropical Diseases1678-91992007-01-0113364065410.1590/S1678-91992007000300007Purification and functional characterization of two fibrinogenolytic enzymes from Bothrops alternatus venomJ. O. CostaC. B. PetricA. HamaguchiM. I. Homsi-BrandeburgoC. Z. OliveiraA. M. SoaresF. OliveiraTwo 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.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992007000300007snake venomBothrops alternatusmetalloproteasesfunctional characterizationfibrinogenolytic activitydefibrinogenation in vivo |
| spellingShingle | J. O. Costa C. B. Petric A. Hamaguchi M. I. Homsi-Brandeburgo C. Z. Oliveira A. M. Soares F. Oliveira Purification and functional characterization of two fibrinogenolytic enzymes from Bothrops alternatus venom Journal of Venomous Animals and Toxins including Tropical Diseases snake venom Bothrops alternatus metalloproteases functional characterization fibrinogenolytic activity defibrinogenation in vivo |
| title | 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_short | 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 |
| topic | snake venom Bothrops alternatus metalloproteases functional characterization fibrinogenolytic activity defibrinogenation in vivo |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992007000300007 |
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