rBaltMIP, a recombinant alpha-type myotoxin inhibitor from Bothrops alternatus (Rhinocerophis alternatus) snake, as a potential candidate to complement the antivenom therapy
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Outros Autores: | , , , , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1016/j.toxicon.2016.10.018 http://hdl.handle.net/11449/162275 |
Resumo: | Phospholipase A(2) inhibitors (PLIs) are important targets in the search and development of new drugs. This study aimed at evaluating the potential of an alpha-type phospholipase A(2) inhibitor from Bothrops alternatus (Rhinocerophis alternatus) snake in its recombinant form (rBaltMIP) to complement the conventional antivenom therapy. Biochemical experiments showed that rBaltMIP presented pl 5.8 and molecular masses of similar to 21 kDa by SDS-PAGE and 19.57 kDa by MALDI/TOF MS. After tryptic peptides sequencing, the results were compared with other PLIs available in databases, showing 100% identity between rBaltMIP and its native inhibitor BaItMIP and from 92% to 96% identity with other inhibitors. Myotoxic activities of BthTX-1 and BthTX-II toxins were measured via plasma CI(levels, showing myotoxic effective concentrations (EC50) of 0.1256 p.g/ 1, and 0.6183 mu g/mu L, respectively. rBaltMIP neutralized the myotoxicity caused by these two toxins up to 65%, without promoting primary antibody response against itself. Nevertheless, this recombinant PLI was immunogenic when standard immunization protocol with Freud's adjuvant was used. In paw edema assays, EC50 of 0.02581 mu g/mu L and 0.02810 mu g/mu L, respectively, were observed with edema reductions of up to 40% by rBaltMIP, suggesting its use as an additional antivenom. In addition, myotoxicity neutralization experiments with the myotoxin BthTX-I showed that rBaltMIP was more effective in inhibiting muscle damage than the conventional anti venom. Thus, considering the severity of envenomations due to Bothrops alternatus (Rhinocerophis alternatus) and the low neutralization of their local effects (such as myotoxicity) by the current anti venoms, rBaltMIP is a promising molecule for the development of novel therapeutic strategies for clinical applications. (C) 2016 Elsevier Ltd. All rights reserved. |
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rBaltMIP, a recombinant alpha-type myotoxin inhibitor from Bothrops alternatus (Rhinocerophis alternatus) snake, as a potential candidate to complement the antivenom therapyBothrops alternatusrBaltMIPPhospholipase A(2) inhibitoralpha PLIPhospholipase A(2) inhibitors (PLIs) are important targets in the search and development of new drugs. This study aimed at evaluating the potential of an alpha-type phospholipase A(2) inhibitor from Bothrops alternatus (Rhinocerophis alternatus) snake in its recombinant form (rBaltMIP) to complement the conventional antivenom therapy. Biochemical experiments showed that rBaltMIP presented pl 5.8 and molecular masses of similar to 21 kDa by SDS-PAGE and 19.57 kDa by MALDI/TOF MS. After tryptic peptides sequencing, the results were compared with other PLIs available in databases, showing 100% identity between rBaltMIP and its native inhibitor BaItMIP and from 92% to 96% identity with other inhibitors. Myotoxic activities of BthTX-1 and BthTX-II toxins were measured via plasma CI(levels, showing myotoxic effective concentrations (EC50) of 0.1256 p.g/ 1, and 0.6183 mu g/mu L, respectively. rBaltMIP neutralized the myotoxicity caused by these two toxins up to 65%, without promoting primary antibody response against itself. Nevertheless, this recombinant PLI was immunogenic when standard immunization protocol with Freud's adjuvant was used. In paw edema assays, EC50 of 0.02581 mu g/mu L and 0.02810 mu g/mu L, respectively, were observed with edema reductions of up to 40% by rBaltMIP, suggesting its use as an additional antivenom. In addition, myotoxicity neutralization experiments with the myotoxin BthTX-I showed that rBaltMIP was more effective in inhibiting muscle damage than the conventional anti venom. Thus, considering the severity of envenomations due to Bothrops alternatus (Rhinocerophis alternatus) and the low neutralization of their local effects (such as myotoxicity) by the current anti venoms, rBaltMIP is a promising molecule for the development of novel therapeutic strategies for clinical applications. (C) 2016 Elsevier Ltd. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Nucleo de Apoio a Pesquisa em Doencas Inflamatorias (NAPDIN)Univ Sao Paulo, FCFRP, Dept Anal Clin Toxicol & Bromatol, Ribeirao Preto, SP, BrazilUniv Fed Sao Carlos, UFScar, Dept Genet & Evolucao, Sao Carlos, SP, BrazilUniv Fed Uberlandia, Inst Ciencias Biomed, Dept Ciencias Fisiol, Uberlandia, MG, BrazilUniv Estadual Paulista, Inst Quim, Dept Bioquim & Tecnol Quim, Araraquara, SP, BrazilUniv Sao Paulo, FMRP, Dept Biol Celular & Mol & Bioagentes Patogen, Ribeirao Preto, SP, BrazilUniv Estadual Paulista, Inst Quim, Dept Bioquim & Tecnol Quim, Araraquara, SP, BrazilFAPESP: 2008/10760-4FAPESP: 2011/23236-4CNPq: 467646/2014-7CNPq: 476932/2012-2Nucleo de Apoio a Pesquisa em Doencas Inflamatorias (NAPDIN): 11.1.21625.01.0Elsevier B.V.Universidade de São Paulo (USP)Universidade Federal de São Carlos (UFSCar)Universidade Federal de Uberlândia (UFU)Universidade Estadual Paulista (Unesp)Santos-Filho, Norival A. [UNESP]Sousa, Tiago S.Boldrini-Franca, JoharaSantos-Silva, Ludier K.Menaldo, Danilo L.Henrique-Silva, FlavioCintra, Adelia C. O.Laure, Helen J.Mamede, Carla C. N.Oliveira, FabioRiul, Thalita B.Dias-Baruffi, MarceloRosa, Jose C.Sampaio, Suely V.2018-11-26T17:15:26Z2018-11-26T17:15:26Z2016-12-15info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article53-62application/pdfhttp://dx.doi.org/10.1016/j.toxicon.2016.10.018Toxicon. Oxford: Pergamon-elsevier Science Ltd, v. 124, p. 53-62, 2016.0041-0101http://hdl.handle.net/11449/16227510.1016/j.toxicon.2016.10.018WOS:000390518600007WOS000390518600007.pdfWeb of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengToxicon0,692info:eu-repo/semantics/openAccess2023-11-13T06:16:16Zoai:repositorio.unesp.br:11449/162275Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T17:36:04.356735Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
rBaltMIP, a recombinant alpha-type myotoxin inhibitor from Bothrops alternatus (Rhinocerophis alternatus) snake, as a potential candidate to complement the antivenom therapy |
title |
rBaltMIP, a recombinant alpha-type myotoxin inhibitor from Bothrops alternatus (Rhinocerophis alternatus) snake, as a potential candidate to complement the antivenom therapy |
spellingShingle |
rBaltMIP, a recombinant alpha-type myotoxin inhibitor from Bothrops alternatus (Rhinocerophis alternatus) snake, as a potential candidate to complement the antivenom therapy Santos-Filho, Norival A. [UNESP] Bothrops alternatus rBaltMIP Phospholipase A(2) inhibitor alpha PLI |
title_short |
rBaltMIP, a recombinant alpha-type myotoxin inhibitor from Bothrops alternatus (Rhinocerophis alternatus) snake, as a potential candidate to complement the antivenom therapy |
title_full |
rBaltMIP, a recombinant alpha-type myotoxin inhibitor from Bothrops alternatus (Rhinocerophis alternatus) snake, as a potential candidate to complement the antivenom therapy |
title_fullStr |
rBaltMIP, a recombinant alpha-type myotoxin inhibitor from Bothrops alternatus (Rhinocerophis alternatus) snake, as a potential candidate to complement the antivenom therapy |
title_full_unstemmed |
rBaltMIP, a recombinant alpha-type myotoxin inhibitor from Bothrops alternatus (Rhinocerophis alternatus) snake, as a potential candidate to complement the antivenom therapy |
title_sort |
rBaltMIP, a recombinant alpha-type myotoxin inhibitor from Bothrops alternatus (Rhinocerophis alternatus) snake, as a potential candidate to complement the antivenom therapy |
author |
Santos-Filho, Norival A. [UNESP] |
author_facet |
Santos-Filho, Norival A. [UNESP] Sousa, Tiago S. Boldrini-Franca, Johara Santos-Silva, Ludier K. Menaldo, Danilo L. Henrique-Silva, Flavio Cintra, Adelia C. O. Laure, Helen J. Mamede, Carla C. N. Oliveira, Fabio Riul, Thalita B. Dias-Baruffi, Marcelo Rosa, Jose C. Sampaio, Suely V. |
author_role |
author |
author2 |
Sousa, Tiago S. Boldrini-Franca, Johara Santos-Silva, Ludier K. Menaldo, Danilo L. Henrique-Silva, Flavio Cintra, Adelia C. O. Laure, Helen J. Mamede, Carla C. N. Oliveira, Fabio Riul, Thalita B. Dias-Baruffi, Marcelo Rosa, Jose C. Sampaio, Suely V. |
author2_role |
author author author author author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade de São Paulo (USP) Universidade Federal de São Carlos (UFSCar) Universidade Federal de Uberlândia (UFU) Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
Santos-Filho, Norival A. [UNESP] Sousa, Tiago S. Boldrini-Franca, Johara Santos-Silva, Ludier K. Menaldo, Danilo L. Henrique-Silva, Flavio Cintra, Adelia C. O. Laure, Helen J. Mamede, Carla C. N. Oliveira, Fabio Riul, Thalita B. Dias-Baruffi, Marcelo Rosa, Jose C. Sampaio, Suely V. |
dc.subject.por.fl_str_mv |
Bothrops alternatus rBaltMIP Phospholipase A(2) inhibitor alpha PLI |
topic |
Bothrops alternatus rBaltMIP Phospholipase A(2) inhibitor alpha PLI |
description |
Phospholipase A(2) inhibitors (PLIs) are important targets in the search and development of new drugs. This study aimed at evaluating the potential of an alpha-type phospholipase A(2) inhibitor from Bothrops alternatus (Rhinocerophis alternatus) snake in its recombinant form (rBaltMIP) to complement the conventional antivenom therapy. Biochemical experiments showed that rBaltMIP presented pl 5.8 and molecular masses of similar to 21 kDa by SDS-PAGE and 19.57 kDa by MALDI/TOF MS. After tryptic peptides sequencing, the results were compared with other PLIs available in databases, showing 100% identity between rBaltMIP and its native inhibitor BaItMIP and from 92% to 96% identity with other inhibitors. Myotoxic activities of BthTX-1 and BthTX-II toxins were measured via plasma CI(levels, showing myotoxic effective concentrations (EC50) of 0.1256 p.g/ 1, and 0.6183 mu g/mu L, respectively. rBaltMIP neutralized the myotoxicity caused by these two toxins up to 65%, without promoting primary antibody response against itself. Nevertheless, this recombinant PLI was immunogenic when standard immunization protocol with Freud's adjuvant was used. In paw edema assays, EC50 of 0.02581 mu g/mu L and 0.02810 mu g/mu L, respectively, were observed with edema reductions of up to 40% by rBaltMIP, suggesting its use as an additional antivenom. In addition, myotoxicity neutralization experiments with the myotoxin BthTX-I showed that rBaltMIP was more effective in inhibiting muscle damage than the conventional anti venom. Thus, considering the severity of envenomations due to Bothrops alternatus (Rhinocerophis alternatus) and the low neutralization of their local effects (such as myotoxicity) by the current anti venoms, rBaltMIP is a promising molecule for the development of novel therapeutic strategies for clinical applications. (C) 2016 Elsevier Ltd. All rights reserved. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-12-15 2018-11-26T17:15:26Z 2018-11-26T17:15:26Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1016/j.toxicon.2016.10.018 Toxicon. Oxford: Pergamon-elsevier Science Ltd, v. 124, p. 53-62, 2016. 0041-0101 http://hdl.handle.net/11449/162275 10.1016/j.toxicon.2016.10.018 WOS:000390518600007 WOS000390518600007.pdf |
url |
http://dx.doi.org/10.1016/j.toxicon.2016.10.018 http://hdl.handle.net/11449/162275 |
identifier_str_mv |
Toxicon. Oxford: Pergamon-elsevier Science Ltd, v. 124, p. 53-62, 2016. 0041-0101 10.1016/j.toxicon.2016.10.018 WOS:000390518600007 WOS000390518600007.pdf |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Toxicon 0,692 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
53-62 application/pdf |
dc.publisher.none.fl_str_mv |
Elsevier B.V. |
publisher.none.fl_str_mv |
Elsevier B.V. |
dc.source.none.fl_str_mv |
Web of Science reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1808128834082439168 |