Molecular identification and activity upon chromogenic substrates of a venombin A from Bothrops atrox Peruvian snake venom
Descripción del Articulo
n this work, the thrombin-like enzyme (TLE) from Bothrops atrox has been identified by mass spectrometry and its enzymatic activity evaluated upon several synthetic substrates. The enzyme was purified to homogeneity using three chromatography steps on Sephadex G-75, CM-Sephadex C-50 and Agarose-PAB....
| Autores: | , , , , |
|---|---|
| Formato: | artículo |
| Fecha de Publicación: | 2010 |
| Institución: | Universidad Nacional Mayor de San Marcos |
| Repositorio: | Revistas - Universidad Nacional Mayor de San Marcos |
| Lenguaje: | español |
| OAI Identifier: | oai:ojs.csi.unmsm:article/12 |
| Enlace del recurso: | https://revistasinvestigacion.unmsm.edu.pe/index.php/rpb/article/view/12 |
| Nivel de acceso: | acceso abierto |
| Materia: | Bothrops atrox enzima similar a trombina MALDI-TOF sustratos cromogénicos. Thrombin-like enzyme MALDI-TOF chromogenic substrates. |
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Revistas - Universidad Nacional Mayor de San Marcos |
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| dc.title.none.fl_str_mv |
Molecular identification and activity upon chromogenic substrates of a venombin A from Bothrops atrox Peruvian snake venom Identificación molecular y actividad sobre sustratos cromogénicos de la venombina A del veneno de la serpiente peruana Bothrops atrox |
| title |
Molecular identification and activity upon chromogenic substrates of a venombin A from Bothrops atrox Peruvian snake venom |
| spellingShingle |
Molecular identification and activity upon chromogenic substrates of a venombin A from Bothrops atrox Peruvian snake venom Sandoval, Gustavo A. Bothrops atrox enzima similar a trombina MALDI-TOF sustratos cromogénicos. Bothrops atrox Thrombin-like enzyme MALDI-TOF chromogenic substrates. |
| title_short |
Molecular identification and activity upon chromogenic substrates of a venombin A from Bothrops atrox Peruvian snake venom |
| title_full |
Molecular identification and activity upon chromogenic substrates of a venombin A from Bothrops atrox Peruvian snake venom |
| title_fullStr |
Molecular identification and activity upon chromogenic substrates of a venombin A from Bothrops atrox Peruvian snake venom |
| title_full_unstemmed |
Molecular identification and activity upon chromogenic substrates of a venombin A from Bothrops atrox Peruvian snake venom |
| title_sort |
Molecular identification and activity upon chromogenic substrates of a venombin A from Bothrops atrox Peruvian snake venom |
| dc.creator.none.fl_str_mv |
Sandoval, Gustavo A. Lazo, Fanny Rodriguez, Edith Yarlequé, Armando Zingali, Russolina B. |
| author |
Sandoval, Gustavo A. |
| author_facet |
Sandoval, Gustavo A. Lazo, Fanny Rodriguez, Edith Yarlequé, Armando Zingali, Russolina B. |
| author_role |
author |
| author2 |
Lazo, Fanny Rodriguez, Edith Yarlequé, Armando Zingali, Russolina B. |
| author2_role |
author author author author |
| dc.subject.none.fl_str_mv |
Bothrops atrox enzima similar a trombina MALDI-TOF sustratos cromogénicos. Bothrops atrox Thrombin-like enzyme MALDI-TOF chromogenic substrates. |
| topic |
Bothrops atrox enzima similar a trombina MALDI-TOF sustratos cromogénicos. Bothrops atrox Thrombin-like enzyme MALDI-TOF chromogenic substrates. |
| description |
n this work, the thrombin-like enzyme (TLE) from Bothrops atrox has been identified by mass spectrometry and its enzymatic activity evaluated upon several synthetic substrates. The enzyme was purified to homogeneity using three chromatography steps on Sephadex G-75, CM-Sephadex C-50 and Agarose-PAB. Also, molecular weight by PAGE-SDS was determined. For molecular identification of this enzyme, mass spectrometry-based peptide mass fingerprinting was used and later in silico analysis. Enzymatic activities were determined using bovine fibrinogen, BApNA and also upon specific chromogenic substrates such as S-2238, S-2251 y S-2266. As a result of these biochemical and structural procedures, we obtained a TLE from B. atrox venom with a molecular weight of 29,6 kDa. Mass spectrometry analysis of obtained peptides, allow us to identify this enzyme as a venombin A, showing a 75% sequence homology. After recording enzymatic activity, this TLE showed coagulant activity on bovine fibrinogen and upon BApNA, S-2238 y S-2266, being unable to hydrolyze S-2251 substrate. Using this combination of structural and functional approaches, we have identified the main component of B. atrox venom related to its coagulant activity, as well as a detailed evaluation of its enzymatic activity upon several substrates |
| publishDate |
2010 |
| dc.date.none.fl_str_mv |
2010-12-31 |
| dc.type.none.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
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article |
| status_str |
publishedVersion |
| dc.identifier.none.fl_str_mv |
https://revistasinvestigacion.unmsm.edu.pe/index.php/rpb/article/view/12 10.15381/rpb.v17i3.12 |
| url |
https://revistasinvestigacion.unmsm.edu.pe/index.php/rpb/article/view/12 |
| identifier_str_mv |
10.15381/rpb.v17i3.12 |
| dc.language.none.fl_str_mv |
spa |
| language |
spa |
| dc.relation.none.fl_str_mv |
https://revistasinvestigacion.unmsm.edu.pe/index.php/rpb/article/view/12/12 |
| dc.rights.none.fl_str_mv |
https://creativecommons.org/licenses/by-nc-sa/4.0 info:eu-repo/semantics/openAccess |
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https://creativecommons.org/licenses/by-nc-sa/4.0 |
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openAccess |
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application/pdf |
| dc.publisher.none.fl_str_mv |
Universidad Nacional Mayor de San Marcos, Facultad de Ciencias Biológicas |
| publisher.none.fl_str_mv |
Universidad Nacional Mayor de San Marcos, Facultad de Ciencias Biológicas |
| dc.source.none.fl_str_mv |
Revista Peruana de Biología; Vol. 17 Núm. 3 (2010); 365 - 370 Revista Peruana de Biología; Vol. 17 No. 3 (2010); 365 - 370 1727-9933 1561-0837 reponame:Revistas - Universidad Nacional Mayor de San Marcos instname:Universidad Nacional Mayor de San Marcos instacron:UNMSM |
| instname_str |
Universidad Nacional Mayor de San Marcos |
| instacron_str |
UNMSM |
| institution |
UNMSM |
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Revistas - Universidad Nacional Mayor de San Marcos |
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Revistas - Universidad Nacional Mayor de San Marcos |
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1795238304242728960 |
| spelling |
Molecular identification and activity upon chromogenic substrates of a venombin A from Bothrops atrox Peruvian snake venomIdentificación molecular y actividad sobre sustratos cromogénicos de la venombina A del veneno de la serpiente peruana Bothrops atroxSandoval, Gustavo A.Lazo, FannyRodriguez, EdithYarlequé, ArmandoZingali, Russolina B.Bothrops atroxenzima similar a trombinaMALDI-TOFsustratos cromogénicos.Bothrops atroxThrombin-like enzyme MALDI-TOFchromogenic substrates.n this work, the thrombin-like enzyme (TLE) from Bothrops atrox has been identified by mass spectrometry and its enzymatic activity evaluated upon several synthetic substrates. The enzyme was purified to homogeneity using three chromatography steps on Sephadex G-75, CM-Sephadex C-50 and Agarose-PAB. Also, molecular weight by PAGE-SDS was determined. For molecular identification of this enzyme, mass spectrometry-based peptide mass fingerprinting was used and later in silico analysis. Enzymatic activities were determined using bovine fibrinogen, BApNA and also upon specific chromogenic substrates such as S-2238, S-2251 y S-2266. As a result of these biochemical and structural procedures, we obtained a TLE from B. atrox venom with a molecular weight of 29,6 kDa. Mass spectrometry analysis of obtained peptides, allow us to identify this enzyme as a venombin A, showing a 75% sequence homology. After recording enzymatic activity, this TLE showed coagulant activity on bovine fibrinogen and upon BApNA, S-2238 y S-2266, being unable to hydrolyze S-2251 substrate. Using this combination of structural and functional approaches, we have identified the main component of B. atrox venom related to its coagulant activity, as well as a detailed evaluation of its enzymatic activity upon several substratesEn el presente trabajo se ha realizado la identificación molecular de la enzima similar a trombina (EST) del veneno de Bothrops atrox y se ha evaluado su actividad enzimática sobre diversos sustratos sintéticos. La enzima fue purificada utilizando tres pasos cromatrográficos, sobre Sephadex G-75, CM-Sephadex C-50 y Agarosa-PAB, determinándose su peso molecular por PAGE-SDS. La identificación molecular de la enzima aislada se realizó por la técnica de peptide mass fingerprinting basada en espectrometría de masas MALDI-TOF y posterior análisis in silico. Las actividades fibrinocoagulante y amidolítica fueron ensayadas sobre fibrinó- geno bovino y BApNA, respectivamente, así como la hidrólisis sobre los sustratos cromogénicos específicos S-2238, S-2251 y S-2266. Como resultado de los ensayos bioquímicos y estructurales, la EST del veneno de B. atrox, presentó un peso molecular de 29,6 kDa. El análisis mediante espectrometría de masas de los péptidos obtenidos, permitió identificar a esta enzima como una venombina A, presentando una identidad del 75%. Del análisis de actividad enzimática, se obtuvo que la EST de B. atrox produjo coagulación del fibrinógeno bovino y presentó actividad sobre BApNA, S-2238 y S-2266, siendo incapaz de hidrolizar el sustrato S-2251. El empleo de estas aproximaciones estructurales y funcionales ha permitido lograr la identificación molecular del principal componente del veneno de B. atrox relacionado con su acción coagulante, así como evaluar en detalle la naturaleza de su actividad enzimática sobre diversos sustratos.Universidad Nacional Mayor de San Marcos, Facultad de Ciencias Biológicas2010-12-31info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://revistasinvestigacion.unmsm.edu.pe/index.php/rpb/article/view/1210.15381/rpb.v17i3.12Revista Peruana de Biología; Vol. 17 Núm. 3 (2010); 365 - 370Revista Peruana de Biología; Vol. 17 No. 3 (2010); 365 - 3701727-99331561-0837reponame:Revistas - Universidad Nacional Mayor de San Marcosinstname:Universidad Nacional Mayor de San Marcosinstacron:UNMSMspahttps://revistasinvestigacion.unmsm.edu.pe/index.php/rpb/article/view/12/12Derechos de autor 2010 Gustavo A. Sandoval, Fanny Lazo, Edith Rodriguez, Armando Yarlequé, Russolina B. Zingalihttps://creativecommons.org/licenses/by-nc-sa/4.0info:eu-repo/semantics/openAccessoai:ojs.csi.unmsm:article/122020-05-21T17:15:10Z |
| score |
13.982926 |
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La información contenida en este registro es de entera responsabilidad de la institución que gestiona el repositorio institucional donde esta contenido este documento o set de datos. El CONCYTEC no se hace responsable por los contenidos (publicaciones y/o datos) accesibles a través del Repositorio Nacional Digital de Ciencia, Tecnología e Innovación de Acceso Abierto (ALICIA).
La información contenida en este registro es de entera responsabilidad de la institución que gestiona el repositorio institucional donde esta contenido este documento o set de datos. El CONCYTEC no se hace responsable por los contenidos (publicaciones y/o datos) accesibles a través del Repositorio Nacional Digital de Ciencia, Tecnología e Innovación de Acceso Abierto (ALICIA).
