Biochemical and biological characterization of two serine proteinases from Colombian Crotalus durissus cumanensis snake venom

Two clotting serine proteinases, named Cdc SI and Cdc SII, were isolated and characterized for the first time from Colombian Crotalus durissus cumanensis snake venom. The enzymes were purified using two chromatographic steps: molecular exclusion on Sephacryl S-200 and RP-HPLC on C8 Column. The molec...

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Detalles Bibliográficos
Autores: Patiño Llano, Arley Camilo, Pereañez, Jaime Andrés, Gutiérrez, José María, Rucavado Romero, Alexandra
Tipo de recurso: artículo
Fecha de publicación:2013
País:Costa Rica
Institución:Universidad de Costa Rica
Repositorio:Kérwá
OAI Identifier:oai:kerwa.ucr.ac.cr:10669/30077
Acceso en línea:http://www.sciencedirect.com/science/article/pii/S0041010112008057
https://hdl.handle.net/10669/30077
Access Level:acceso embargado
Palabra clave:Thrombin-like enzyme
Coagulant activity
Crotalus durissus cumanensis
Serine proteinase
Cdc SI
Cdc SII
Snake venom
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dc.title.es_ES.fl_str_mv Biochemical and biological characterization of two serine proteinases from Colombian Crotalus durissus cumanensis snake venom
title Biochemical and biological characterization of two serine proteinases from Colombian Crotalus durissus cumanensis snake venom
spellingShingle Biochemical and biological characterization of two serine proteinases from Colombian Crotalus durissus cumanensis snake venom
Patiño Llano, Arley Camilo
Thrombin-like enzyme
Coagulant activity
Crotalus durissus cumanensis
Serine proteinase
Cdc SI
Cdc SII
Snake venom
title_short Biochemical and biological characterization of two serine proteinases from Colombian Crotalus durissus cumanensis snake venom
title_full Biochemical and biological characterization of two serine proteinases from Colombian Crotalus durissus cumanensis snake venom
title_fullStr Biochemical and biological characterization of two serine proteinases from Colombian Crotalus durissus cumanensis snake venom
title_full_unstemmed Biochemical and biological characterization of two serine proteinases from Colombian Crotalus durissus cumanensis snake venom
title_sort Biochemical and biological characterization of two serine proteinases from Colombian Crotalus durissus cumanensis snake venom
dc.creator.none.fl_str_mv Patiño Llano, Arley Camilo
Pereañez, Jaime Andrés
Gutiérrez, José María
Rucavado Romero, Alexandra
author Patiño Llano, Arley Camilo
author_facet Patiño Llano, Arley Camilo
Pereañez, Jaime Andrés
Gutiérrez, José María
Rucavado Romero, Alexandra
author_role author
author2 Pereañez, Jaime Andrés
Gutiérrez, José María
Rucavado Romero, Alexandra
author2_role author
author
author
dc.subject.es_ES.fl_str_mv Thrombin-like enzyme
Coagulant activity
Crotalus durissus cumanensis
Serine proteinase
Cdc SI
Cdc SII
Snake venom
topic Thrombin-like enzyme
Coagulant activity
Crotalus durissus cumanensis
Serine proteinase
Cdc SI
Cdc SII
Snake venom
description Two clotting serine proteinases, named Cdc SI and Cdc SII, were isolated and characterized for the first time from Colombian Crotalus durissus cumanensis snake venom. The enzymes were purified using two chromatographic steps: molecular exclusion on Sephacryl S-200 and RP-HPLC on C8 Column. The molecular masses of the proteins, determined by MALDI-TOF mass spectrometry, were 28,561.4 and 28,799.2 Da for Cdc SI and Cdc SII, respectively. The aim of the present study was to evaluate enzymatic, coagulant and toxic properties of the two enzymes. The serine proteinases hydrolyzed specific chromogenic substrate (BaPNA) and exhibited a Michaelis–Menten behavior. Cdc SI had Vmax of 0.038 ± 0.003 nmol/min and KM of 0.034 ± 0.017 mM, while Cdc SII displayed values of Vmax of 0.267 ± 0.011 nmol/min and KM of 0.145 ± 0.023 mM. N-terminal sequences were VIGGDEXNIN and VIGGDICNINEHNFLVALYE for Cdc SI and Cdc SII, respectively. Molecular masses, N-terminal sequences, inhibition assays, and enzymatic profile suggest that Cdc SI and Cdc SII belong to the family of snake venom thrombin-like enzymes. These serine proteinases differed in their clotting activity on human plasma, showing a minimum coagulant dose of 25 μg and 0.571 μg for Cdc SI and Cdc SII, respectively. Enzymes also showed coagulant activity on bovine fibrinogen and degraded chain α of this protein. Toxins lack hemorrhagic and myotoxic activities, but are capable to induce defibrin(ogen)ation, moderate edema, and an increase in vascular permeability. These serine proteinases may contribute indirectly to the local hemorrhage induced by metalloproteinases, by causing blood clotting disturbances, and might also contribute to cardiovascular alterations characteristic of patients envenomed by C. d. cumanensis in Colombia.
publishDate 2013
dc.date.issued.none.fl_str_mv 2013-03-01
dc.date.accessioned.none.fl_str_mv 2017-06-09T14:05:21Z
dc.date.available.none.fl_str_mv 2017-06-09T14:05:21Z
dc.type.none.fl_str_mv artículo original
http://purl.org/coar/resource_type/c_2df8fbb1
info:eu-repo/semantics/article
format article
dc.identifier.citation.none.fl_str_mv http://www.sciencedirect.com/science/article/pii/S0041010112008057
dc.identifier.issn.none.fl_str_mv 0041-0101
dc.identifier.uri.none.fl_str_mv https://hdl.handle.net/10669/30077
dc.identifier.doi.none.fl_str_mv 10.1016/j.toxicon.2012.11.010
dc.identifier.pmid.none.fl_str_mv 23178323
url http://www.sciencedirect.com/science/article/pii/S0041010112008057
https://hdl.handle.net/10669/30077
identifier_str_mv 0041-0101
10.1016/j.toxicon.2012.11.010
23178323
dc.language.iso.es_ES.fl_str_mv en_US
language_invalid_str_mv en_US
dc.rights.none.fl_str_mv acceso embargado
http://purl.org/coar/access_right/c_f1cf
info:eu-repo/semantics/embargoedAccess
rights_invalid_str_mv acceso embargado
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eu_rights_str_mv embargoedAccess
dc.source.es_ES.fl_str_mv Toxicon; Volumen 63. 2013
dc.source.none.fl_str_mv reponame:Kérwá
instname:Universidad de Costa Rica
instacron:UCR
instname_str Universidad de Costa Rica
instacron_str UCR
institution UCR
reponame_str Kérwá
collection Kérwá
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spelling Patiño Llano, Arley Camilo7ad1862e-589c-46cb-8e33-9100a654c71a600Pereañez, Jaime Andrésf92ca8a9-0129-44df-9994-a81ea01778f3600Gutiérrez, José María6a9bac5f-130f-4977-8ddf-ab97fcafacc4600Rucavado Romero, Alexandraf62bf3bf-c612-451f-90ef-a366ebb3f4b56002017-06-09T14:05:21Z2017-06-09T14:05:21Z2013-03-01http://www.sciencedirect.com/science/article/pii/S00410101120080570041-0101https://hdl.handle.net/10669/3007710.1016/j.toxicon.2012.11.01023178323Two clotting serine proteinases, named Cdc SI and Cdc SII, were isolated and characterized for the first time from Colombian Crotalus durissus cumanensis snake venom. The enzymes were purified using two chromatographic steps: molecular exclusion on Sephacryl S-200 and RP-HPLC on C8 Column. The molecular masses of the proteins, determined by MALDI-TOF mass spectrometry, were 28,561.4 and 28,799.2 Da for Cdc SI and Cdc SII, respectively. The aim of the present study was to evaluate enzymatic, coagulant and toxic properties of the two enzymes. The serine proteinases hydrolyzed specific chromogenic substrate (BaPNA) and exhibited a Michaelis–Menten behavior. Cdc SI had Vmax of 0.038 ± 0.003 nmol/min and KM of 0.034 ± 0.017 mM, while Cdc SII displayed values of Vmax of 0.267 ± 0.011 nmol/min and KM of 0.145 ± 0.023 mM. N-terminal sequences were VIGGDEXNIN and VIGGDICNINEHNFLVALYE for Cdc SI and Cdc SII, respectively. Molecular masses, N-terminal sequences, inhibition assays, and enzymatic profile suggest that Cdc SI and Cdc SII belong to the family of snake venom thrombin-like enzymes. These serine proteinases differed in their clotting activity on human plasma, showing a minimum coagulant dose of 25 μg and 0.571 μg for Cdc SI and Cdc SII, respectively. Enzymes also showed coagulant activity on bovine fibrinogen and degraded chain α of this protein. Toxins lack hemorrhagic and myotoxic activities, but are capable to induce defibrin(ogen)ation, moderate edema, and an increase in vascular permeability. These serine proteinases may contribute indirectly to the local hemorrhage induced by metalloproteinases, by causing blood clotting disturbances, and might also contribute to cardiovascular alterations characteristic of patients envenomed by C. d. cumanensis in Colombia.Universidad de Costa Rica/[741-B0-506]/UCR/Costa RicaConsejo Nacional de Rectores//CONARE/Costa RicaComité para el desarrollo de la investigación//CODI/ColombiaUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Instituto Clodomiro Picado (ICP)en_USacceso embargadohttp://purl.org/coar/access_right/c_f1cfinfo:eu-repo/semantics/embargoedAccessToxicon; Volumen 63. 2013reponame:Kérwáinstname:Universidad de Costa Ricainstacron:UCRThrombin-like enzymeCoagulant activityCrotalus durissus cumanensisSerine proteinaseCdc SICdc SIISnake venomBiochemical and biological characterization of two serine proteinases from Colombian Crotalus durissus cumanensis snake venomartículo originalhttp://purl.org/coar/resource_type/c_2df8fbb1info:eu-repo/semantics/articleTHUMBNAIL347_2013_Toxicon_Patiño_Crotalus_serine_proteinases.pdf.jpg347_2013_Toxicon_Patiño_Crotalus_serine_proteinases.pdf.jpgGenerated Thumbnailimage/jpeg3366https://www.kerwa.ucr.ac.cr/bitstreams/09669150-ce82-4b51-848a-0a9e05dad617/downloadf2e8d31a03485b015c09151605e53a63MD54ORIGINAL347_2013_Toxicon_Patiño_Crotalus_serine_proteinases.pdf347_2013_Toxicon_Patiño_Crotalus_serine_proteinases.pdfVersión finalapplication/pdf1343219https://www.kerwa.ucr.ac.cr/bitstreams/64fe5e8b-1680-4204-a14d-395f06f7125b/download27b0507b99fba8b0ae974c5ade1f68a9MD51LICENSElicense.txtlicense.txttext/plain; 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