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Rapid Purification and Procoagulant and Platelet Aggregating Activities of Rhombeobin: A Thrombin-Like/Gyroxin-Like Enzyme from Lachesis muta rhombeata Snake Venom

We report a rapid purification method using one-step chromatography of SVSP Rhombeobin (LMR-47) from Lachesis muta rhombeata venom and its procoagulant activities and effects on platelet aggregation. The venom was fractionated by a single chromatographic step in RP-HPLC on a C8 Discovery BIO Wide Po...

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Autores principales: Torres-Huaco, Frank Denis, Werneck, Cláudio C., Vicente, Cristina Pontes, Vassequi-Silva, Talita, Nery-Diez, Ana Cláudia Coelho, Mendes, Camila B., Antunes, Edson, Marangoni, Sérgio, Damico, Daniela C. S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3766598/
https://www.ncbi.nlm.nih.gov/pubmed/24058917
http://dx.doi.org/10.1155/2013/903292
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author Torres-Huaco, Frank Denis
Werneck, Cláudio C.
Vicente, Cristina Pontes
Vassequi-Silva, Talita
Nery-Diez, Ana Cláudia Coelho
Mendes, Camila B.
Antunes, Edson
Marangoni, Sérgio
Damico, Daniela C. S.
author_facet Torres-Huaco, Frank Denis
Werneck, Cláudio C.
Vicente, Cristina Pontes
Vassequi-Silva, Talita
Nery-Diez, Ana Cláudia Coelho
Mendes, Camila B.
Antunes, Edson
Marangoni, Sérgio
Damico, Daniela C. S.
author_sort Torres-Huaco, Frank Denis
collection PubMed
description We report a rapid purification method using one-step chromatography of SVSP Rhombeobin (LMR-47) from Lachesis muta rhombeata venom and its procoagulant activities and effects on platelet aggregation. The venom was fractionated by a single chromatographic step in RP-HPLC on a C8 Discovery BIO Wide Pore, showing high degree of molecular homogeneity with molecular mass of 47035.49 Da. Rhombeobin showed amidolytic activity upon BAρNA, with a broad optimum pH (7–10) and was stable in solution up to 60°C. The amidolytic activity was inhibited by serine proteinase inhibitors and reducing agents, but not chelating agents. Rhombeobin showed high coagulant activity on mice plasma and bovine fibrinogen. The deduced amino acid sequence of Rhombeobin showed homology with other SVSPs, especially with LM-TL (L. m. muta) and Gyroxin (C. d. terrificus). Rhombeobin acts, in vitro, as a strong procoagulant enzyme on mice citrated plasma, shortening the APTT and PT tests in adose-dependent manner. The protein showed, “ex vivo”, a strong defibrinogenating effect with 1 µg/animal. Lower doses activated the intrinsic and extrinsic coagulation pathways and impaired the platelet aggregation induced by ADP. Thus, this is the first report of a venom component that produces a venom-induced consumptive coagulopathy (VICC).
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spelling pubmed-37665982013-09-22 Rapid Purification and Procoagulant and Platelet Aggregating Activities of Rhombeobin: A Thrombin-Like/Gyroxin-Like Enzyme from Lachesis muta rhombeata Snake Venom Torres-Huaco, Frank Denis Werneck, Cláudio C. Vicente, Cristina Pontes Vassequi-Silva, Talita Nery-Diez, Ana Cláudia Coelho Mendes, Camila B. Antunes, Edson Marangoni, Sérgio Damico, Daniela C. S. Biomed Res Int Research Article We report a rapid purification method using one-step chromatography of SVSP Rhombeobin (LMR-47) from Lachesis muta rhombeata venom and its procoagulant activities and effects on platelet aggregation. The venom was fractionated by a single chromatographic step in RP-HPLC on a C8 Discovery BIO Wide Pore, showing high degree of molecular homogeneity with molecular mass of 47035.49 Da. Rhombeobin showed amidolytic activity upon BAρNA, with a broad optimum pH (7–10) and was stable in solution up to 60°C. The amidolytic activity was inhibited by serine proteinase inhibitors and reducing agents, but not chelating agents. Rhombeobin showed high coagulant activity on mice plasma and bovine fibrinogen. The deduced amino acid sequence of Rhombeobin showed homology with other SVSPs, especially with LM-TL (L. m. muta) and Gyroxin (C. d. terrificus). Rhombeobin acts, in vitro, as a strong procoagulant enzyme on mice citrated plasma, shortening the APTT and PT tests in adose-dependent manner. The protein showed, “ex vivo”, a strong defibrinogenating effect with 1 µg/animal. Lower doses activated the intrinsic and extrinsic coagulation pathways and impaired the platelet aggregation induced by ADP. Thus, this is the first report of a venom component that produces a venom-induced consumptive coagulopathy (VICC). Hindawi Publishing Corporation 2013 2013-08-24 /pmc/articles/PMC3766598/ /pubmed/24058917 http://dx.doi.org/10.1155/2013/903292 Text en Copyright © 2013 Frank Denis Torres-Huaco et al. https://creativecommons.org/licenses/by/3.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Torres-Huaco, Frank Denis
Werneck, Cláudio C.
Vicente, Cristina Pontes
Vassequi-Silva, Talita
Nery-Diez, Ana Cláudia Coelho
Mendes, Camila B.
Antunes, Edson
Marangoni, Sérgio
Damico, Daniela C. S.
Rapid Purification and Procoagulant and Platelet Aggregating Activities of Rhombeobin: A Thrombin-Like/Gyroxin-Like Enzyme from Lachesis muta rhombeata Snake Venom
title Rapid Purification and Procoagulant and Platelet Aggregating Activities of Rhombeobin: A Thrombin-Like/Gyroxin-Like Enzyme from Lachesis muta rhombeata Snake Venom
title_full Rapid Purification and Procoagulant and Platelet Aggregating Activities of Rhombeobin: A Thrombin-Like/Gyroxin-Like Enzyme from Lachesis muta rhombeata Snake Venom
title_fullStr Rapid Purification and Procoagulant and Platelet Aggregating Activities of Rhombeobin: A Thrombin-Like/Gyroxin-Like Enzyme from Lachesis muta rhombeata Snake Venom
title_full_unstemmed Rapid Purification and Procoagulant and Platelet Aggregating Activities of Rhombeobin: A Thrombin-Like/Gyroxin-Like Enzyme from Lachesis muta rhombeata Snake Venom
title_short Rapid Purification and Procoagulant and Platelet Aggregating Activities of Rhombeobin: A Thrombin-Like/Gyroxin-Like Enzyme from Lachesis muta rhombeata Snake Venom
title_sort rapid purification and procoagulant and platelet aggregating activities of rhombeobin: a thrombin-like/gyroxin-like enzyme from lachesis muta rhombeata snake venom
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3766598/
https://www.ncbi.nlm.nih.gov/pubmed/24058917
http://dx.doi.org/10.1155/2013/903292
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