Boa.PLI: Structural and functional characterization of the gamma phospholipase A2 plasma inhibitor from the non-venomous Brazilian snake Boa constrictor
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Outros Autores: | , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1371/journal.pone.0229657 http://hdl.handle.net/11449/196909 |
Resumo: | Plasma in several organisms has components that promote resistance to envenomation by inhibiting specific proteins from snake venoms, such as phospholipases A(2) (PLA(2)s). The major hypothesis for inhibitor's presence would be the protection against self-envenomation in venomous snakes, but the occurrence of inhibitors in non-venomous snakes and other animals has opened new perspectives for this molecule. Thus, this study showed for the first time the structural and functional characterization of the PLA(2) inhibitor from the Boa constrictor serum (Boa.PLI), a non-venomous snake that dwells extensively the Brazilian territory. Therefore, the inhibitor was isolated from B. constrictor serum, with 0.63% of recovery. SDS-PAGE showed a band at similar to 25 kDa under reducing conditions and similar to 20 kDa under non-reducing conditions. Chromatographic analyses showed the presence of oligomers formed by Boa.PLI. Primary structure of Boa.PLI suggested an estimated molecular mass of 22 kDa. When Boa.PLI was incubated with Asp-49 and Lys-49 PLA(2) there was no severe change in its dichroism spectrum, suggesting a non-covalent interaction. The enzymatic assay showed a dose-dependent inhibition, up to 48.2%, when Boa.PLI was incubated with Asp-49 PLA(2), since Lys-49 PLA(2) has a lack of enzymatic activity. The edematogenic and myotoxic effects of PLA(2)s were also inhibited by Boa.PLI. In summary, the present work provides new insights into inhibitors from non-venomous snakes, which possess PLIs in their plasma, although the contact with venom is unlikely. |
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Boa.PLI: Structural and functional characterization of the gamma phospholipase A2 plasma inhibitor from the non-venomous Brazilian snake Boa constrictorPlasma in several organisms has components that promote resistance to envenomation by inhibiting specific proteins from snake venoms, such as phospholipases A(2) (PLA(2)s). The major hypothesis for inhibitor's presence would be the protection against self-envenomation in venomous snakes, but the occurrence of inhibitors in non-venomous snakes and other animals has opened new perspectives for this molecule. Thus, this study showed for the first time the structural and functional characterization of the PLA(2) inhibitor from the Boa constrictor serum (Boa.PLI), a non-venomous snake that dwells extensively the Brazilian territory. Therefore, the inhibitor was isolated from B. constrictor serum, with 0.63% of recovery. SDS-PAGE showed a band at similar to 25 kDa under reducing conditions and similar to 20 kDa under non-reducing conditions. Chromatographic analyses showed the presence of oligomers formed by Boa.PLI. Primary structure of Boa.PLI suggested an estimated molecular mass of 22 kDa. When Boa.PLI was incubated with Asp-49 and Lys-49 PLA(2) there was no severe change in its dichroism spectrum, suggesting a non-covalent interaction. The enzymatic assay showed a dose-dependent inhibition, up to 48.2%, when Boa.PLI was incubated with Asp-49 PLA(2), since Lys-49 PLA(2) has a lack of enzymatic activity. The edematogenic and myotoxic effects of PLA(2)s were also inhibited by Boa.PLI. In summary, the present work provides new insights into inhibitors from non-venomous snakes, which possess PLIs in their plasma, although the contact with venom is unlikely.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Univ Sao Paulo, Inst Pesquisas Tecnol, Inst Butantan, Interunidades Biotecnol, Sao Paulo, SP, BrazilInst Butantan, Lab Herpetol, Sao Paulo, SP, BrazilUniv Fed Minas Gerais, Belo Horizonte, MG, BrazilUniv Fed Sao Paulo, Dept Bioquim, Sao Paulo, SP, BrazilUniv Estadual Paulista, Inst Biociencias Litoral Paulista, Sao Vicente, SP, BrazilUniv Estadual Paulista, Inst Biociencias Litoral Paulista, Sao Vicente, SP, BrazilCAPES: 001FAPESP: 2017/16908-2FAPESP: 2017/20106-9FAPESP: 2018/25786-0Public Library ScienceUniversidade de São Paulo (USP)Inst ButantanUniversidade Federal de Minas Gerais (UFMG)Universidade Federal de São Paulo (UNIFESP)Universidade Estadual Paulista (Unesp)Bittencourt Rodrigues, Caroline FabriSerino-Silva, CarolineMorais-Zani, Karen deKavazoi, Victor KoitiNogueira Carvalho, Marcelo PiresGrego, Kathleen FernandesChiarelli, TassiaTashima, Alexandre KeijiToyama, Marcos Hikari [UNESP]Tanaka-Azevedo, Anita Mitico2020-12-10T20:00:06Z2020-12-10T20:00:06Z2020-02-27info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article13http://dx.doi.org/10.1371/journal.pone.0229657Plos One. San Francisco: Public Library Science, v. 15, n. 2, 13 p., 2020.1932-6203http://hdl.handle.net/11449/19690910.1371/journal.pone.0229657WOS:000535237000056Web of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengPlos Oneinfo:eu-repo/semantics/openAccess2021-10-23T10:11:07Zoai:repositorio.unesp.br:11449/196909Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T23:16:40.992148Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Boa.PLI: Structural and functional characterization of the gamma phospholipase A2 plasma inhibitor from the non-venomous Brazilian snake Boa constrictor |
title |
Boa.PLI: Structural and functional characterization of the gamma phospholipase A2 plasma inhibitor from the non-venomous Brazilian snake Boa constrictor |
spellingShingle |
Boa.PLI: Structural and functional characterization of the gamma phospholipase A2 plasma inhibitor from the non-venomous Brazilian snake Boa constrictor Bittencourt Rodrigues, Caroline Fabri |
title_short |
Boa.PLI: Structural and functional characterization of the gamma phospholipase A2 plasma inhibitor from the non-venomous Brazilian snake Boa constrictor |
title_full |
Boa.PLI: Structural and functional characterization of the gamma phospholipase A2 plasma inhibitor from the non-venomous Brazilian snake Boa constrictor |
title_fullStr |
Boa.PLI: Structural and functional characterization of the gamma phospholipase A2 plasma inhibitor from the non-venomous Brazilian snake Boa constrictor |
title_full_unstemmed |
Boa.PLI: Structural and functional characterization of the gamma phospholipase A2 plasma inhibitor from the non-venomous Brazilian snake Boa constrictor |
title_sort |
Boa.PLI: Structural and functional characterization of the gamma phospholipase A2 plasma inhibitor from the non-venomous Brazilian snake Boa constrictor |
author |
Bittencourt Rodrigues, Caroline Fabri |
author_facet |
Bittencourt Rodrigues, Caroline Fabri Serino-Silva, Caroline Morais-Zani, Karen de Kavazoi, Victor Koiti Nogueira Carvalho, Marcelo Pires Grego, Kathleen Fernandes Chiarelli, Tassia Tashima, Alexandre Keiji Toyama, Marcos Hikari [UNESP] Tanaka-Azevedo, Anita Mitico |
author_role |
author |
author2 |
Serino-Silva, Caroline Morais-Zani, Karen de Kavazoi, Victor Koiti Nogueira Carvalho, Marcelo Pires Grego, Kathleen Fernandes Chiarelli, Tassia Tashima, Alexandre Keiji Toyama, Marcos Hikari [UNESP] Tanaka-Azevedo, Anita Mitico |
author2_role |
author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade de São Paulo (USP) Inst Butantan Universidade Federal de Minas Gerais (UFMG) Universidade Federal de São Paulo (UNIFESP) Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
Bittencourt Rodrigues, Caroline Fabri Serino-Silva, Caroline Morais-Zani, Karen de Kavazoi, Victor Koiti Nogueira Carvalho, Marcelo Pires Grego, Kathleen Fernandes Chiarelli, Tassia Tashima, Alexandre Keiji Toyama, Marcos Hikari [UNESP] Tanaka-Azevedo, Anita Mitico |
description |
Plasma in several organisms has components that promote resistance to envenomation by inhibiting specific proteins from snake venoms, such as phospholipases A(2) (PLA(2)s). The major hypothesis for inhibitor's presence would be the protection against self-envenomation in venomous snakes, but the occurrence of inhibitors in non-venomous snakes and other animals has opened new perspectives for this molecule. Thus, this study showed for the first time the structural and functional characterization of the PLA(2) inhibitor from the Boa constrictor serum (Boa.PLI), a non-venomous snake that dwells extensively the Brazilian territory. Therefore, the inhibitor was isolated from B. constrictor serum, with 0.63% of recovery. SDS-PAGE showed a band at similar to 25 kDa under reducing conditions and similar to 20 kDa under non-reducing conditions. Chromatographic analyses showed the presence of oligomers formed by Boa.PLI. Primary structure of Boa.PLI suggested an estimated molecular mass of 22 kDa. When Boa.PLI was incubated with Asp-49 and Lys-49 PLA(2) there was no severe change in its dichroism spectrum, suggesting a non-covalent interaction. The enzymatic assay showed a dose-dependent inhibition, up to 48.2%, when Boa.PLI was incubated with Asp-49 PLA(2), since Lys-49 PLA(2) has a lack of enzymatic activity. The edematogenic and myotoxic effects of PLA(2)s were also inhibited by Boa.PLI. In summary, the present work provides new insights into inhibitors from non-venomous snakes, which possess PLIs in their plasma, although the contact with venom is unlikely. |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020-12-10T20:00:06Z 2020-12-10T20:00:06Z 2020-02-27 |
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/10.1371/journal.pone.0229657 Plos One. San Francisco: Public Library Science, v. 15, n. 2, 13 p., 2020. 1932-6203 http://hdl.handle.net/11449/196909 10.1371/journal.pone.0229657 WOS:000535237000056 |
url |
http://dx.doi.org/10.1371/journal.pone.0229657 http://hdl.handle.net/11449/196909 |
identifier_str_mv |
Plos One. San Francisco: Public Library Science, v. 15, n. 2, 13 p., 2020. 1932-6203 10.1371/journal.pone.0229657 WOS:000535237000056 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Plos One |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
13 |
dc.publisher.none.fl_str_mv |
Public Library Science |
publisher.none.fl_str_mv |
Public Library Science |
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 |
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1808129503665324032 |