Isolation of bothrasperin, a disintegrin with potent platelet aggregation inhibitory activity, from the venom of the snake Bothrops asper
The venom of Bothrops asper induces severe coagulation disturbances in accidentally envenomed humans. However, only few studies have been conducted to identify components that interact with the hemostatic system in this venom. In the present work, we fractionated B. asper venom in order to investiga...
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Vicerractoría Investigación
2003-03-01
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| Series: | Revista de Biología Tropical |
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| Online Access: | http://www.scielo.sa.cr/scielo.php?script=sci_arttext&pid=S0034-77442003000100024 |
| _version_ | 1797705926440910848 |
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| author | Adrián Pinto Yamileth Angulo Rafael Jiménez Bruno Lomonte |
| author_facet | Adrián Pinto Yamileth Angulo Rafael Jiménez Bruno Lomonte |
| author_sort | Adrián Pinto |
| collection | DOAJ |
| description | The venom of Bothrops asper induces severe coagulation disturbances in accidentally envenomed humans. However, only few studies have been conducted to identify components that interact with the hemostatic system in this venom. In the present work, we fractionated B. asper venom in order to investigate the possible presence of inhibitors of platelet aggregation. Using a combination of gel filtration, anion-exchange chromatography, and reverse-phase high performance liquid chromatography, we isolated an acidic protein which shows a single chain composition, with a molecular mass of ~8 kDa, estimated by SDS-polyacrylamide gel electrophoresis. Its N-terminal sequence has high similarity to disintegrins isolated from different snake venoms, which are known to bind to cellular integrins such as the GPIIb/IIIa fibrinogen receptor on platelets. The purified protein exerted potent aggregation inhibitory activity on ADP-stimulated human platelets in vitro, with an estimated IC50 of 50 nM. This biological activity, together with the biochemical characteristics observed, demonstrate that the protein isolated from B. asper venom is a disintegrin, hereby named "bothrasperin". This is the first disintegrin isolated from Central American viperid snake species.<br>El veneno de la serpiente Bothrops asper induce graves alteraciones de la coagulación en los humanos accidentalmente envenenados. Sin embargo, se han realizado pocos estudios para identificar los componentes del veneno que interactúan con el sistema hemostático. En el presente trabajo, fraccionamos el veneno de B. asper para investigar la posible presencia de inhibidores de la agregación plaquetaria. Empleando una combinación de técnicas cromatográficas (filtración en gel, intercambio aniónico y cromatografía líquida de alto desempeño en fase reversa), aislamos una proteína acídica de cadena simple, con una masa molecular de ~8 kDa, estimada mediante electroforesis en gel de poliacrilamida con SDS. Su secuencia de aminoácidos N-terminal muestra una alta similitud con la de disintegrinas aisladas de diferentes venenos de serpiente, las cuales se unen a integrinas celulares como el receptor de fibrinógeno GPIIb/IIIa de las plaquetas. La proteína purificada ejerce una potente acción inhibitoria sobre la agregación in vitro de plaquetas humanas estimuladas con ADP, con una IC 50 estimada en 50 nM. Esta actividad biológica, sumada a las características bioquímicas observadas, demuestran que la proteína aislada del veneno de B. asper es una disintegrina, a la cual denominamos "both-rasperina". Esta es la primera disintegrina aislada de una especie de vipéridos de Centroamérica. |
| first_indexed | 2024-03-12T05:43:34Z |
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| issn | 0034-7744 2215-2075 |
| language | English |
| last_indexed | 2024-03-12T05:43:34Z |
| publishDate | 2003-03-01 |
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| series | Revista de Biología Tropical |
| spelling | doaj.art-84e34a3f2002438bb4bfaef2aff34d312023-09-03T05:47:42ZengVicerractoría InvestigaciónRevista de Biología Tropical0034-77442215-20752003-03-01511253260Isolation of bothrasperin, a disintegrin with potent platelet aggregation inhibitory activity, from the venom of the snake Bothrops asperAdrián PintoYamileth AnguloRafael JiménezBruno LomonteThe venom of Bothrops asper induces severe coagulation disturbances in accidentally envenomed humans. However, only few studies have been conducted to identify components that interact with the hemostatic system in this venom. In the present work, we fractionated B. asper venom in order to investigate the possible presence of inhibitors of platelet aggregation. Using a combination of gel filtration, anion-exchange chromatography, and reverse-phase high performance liquid chromatography, we isolated an acidic protein which shows a single chain composition, with a molecular mass of ~8 kDa, estimated by SDS-polyacrylamide gel electrophoresis. Its N-terminal sequence has high similarity to disintegrins isolated from different snake venoms, which are known to bind to cellular integrins such as the GPIIb/IIIa fibrinogen receptor on platelets. The purified protein exerted potent aggregation inhibitory activity on ADP-stimulated human platelets in vitro, with an estimated IC50 of 50 nM. This biological activity, together with the biochemical characteristics observed, demonstrate that the protein isolated from B. asper venom is a disintegrin, hereby named "bothrasperin". This is the first disintegrin isolated from Central American viperid snake species.<br>El veneno de la serpiente Bothrops asper induce graves alteraciones de la coagulación en los humanos accidentalmente envenenados. Sin embargo, se han realizado pocos estudios para identificar los componentes del veneno que interactúan con el sistema hemostático. En el presente trabajo, fraccionamos el veneno de B. asper para investigar la posible presencia de inhibidores de la agregación plaquetaria. Empleando una combinación de técnicas cromatográficas (filtración en gel, intercambio aniónico y cromatografía líquida de alto desempeño en fase reversa), aislamos una proteína acídica de cadena simple, con una masa molecular de ~8 kDa, estimada mediante electroforesis en gel de poliacrilamida con SDS. Su secuencia de aminoácidos N-terminal muestra una alta similitud con la de disintegrinas aisladas de diferentes venenos de serpiente, las cuales se unen a integrinas celulares como el receptor de fibrinógeno GPIIb/IIIa de las plaquetas. La proteína purificada ejerce una potente acción inhibitoria sobre la agregación in vitro de plaquetas humanas estimuladas con ADP, con una IC 50 estimada en 50 nM. Esta actividad biológica, sumada a las características bioquímicas observadas, demuestran que la proteína aislada del veneno de B. asper es una disintegrina, a la cual denominamos "both-rasperina". Esta es la primera disintegrina aislada de una especie de vipéridos de Centroamérica.http://www.scielo.sa.cr/scielo.php?script=sci_arttext&pid=S0034-77442003000100024Snake venomdisintegrinBothropsplatelet aggregation |
| spellingShingle | Adrián Pinto Yamileth Angulo Rafael Jiménez Bruno Lomonte Isolation of bothrasperin, a disintegrin with potent platelet aggregation inhibitory activity, from the venom of the snake Bothrops asper Revista de Biología Tropical Snake venom disintegrin Bothrops platelet aggregation |
| title | Isolation of bothrasperin, a disintegrin with potent platelet aggregation inhibitory activity, from the venom of the snake Bothrops asper |
| title_full | Isolation of bothrasperin, a disintegrin with potent platelet aggregation inhibitory activity, from the venom of the snake Bothrops asper |
| title_fullStr | Isolation of bothrasperin, a disintegrin with potent platelet aggregation inhibitory activity, from the venom of the snake Bothrops asper |
| title_full_unstemmed | Isolation of bothrasperin, a disintegrin with potent platelet aggregation inhibitory activity, from the venom of the snake Bothrops asper |
| title_short | Isolation of bothrasperin, a disintegrin with potent platelet aggregation inhibitory activity, from the venom of the snake Bothrops asper |
| title_sort | isolation of bothrasperin a disintegrin with potent platelet aggregation inhibitory activity from the venom of the snake bothrops asper |
| topic | Snake venom disintegrin Bothrops platelet aggregation |
| url | http://www.scielo.sa.cr/scielo.php?script=sci_arttext&pid=S0034-77442003000100024 |
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