Brown spiders’ phospholipases-d with potential therapeutic applications: Functional assessment of mutant isoforms
Autor(a) principal: | |
---|---|
Data de Publicação: | 2021 |
Outros Autores: | , , , , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.3390/biomedicines9030320 http://hdl.handle.net/11449/208589 |
Resumo: | Phospholipases-D (PLDs) found in Loxosceles spiders’ venoms are responsible for the der-monecrosis triggered by envenomation. PLDs can also induce other local and systemic effects, such as massive inflammatory response, edema, and hemolysis. Recombinant PLDs reproduce all of the deleterious effects induced by Loxosceles whole venoms. Herein, wild type and mutant PLDs of two species involved in accidents—L. gaucho and L. laeta—were recombinantly expressed and character-ized. The mutations are related to amino acid residues relevant for catalysis (H12-H47), magnesium ion coordination (E32-D34) and binding to phospholipid substrates (Y228 and Y228-Y229-W230). Circular dichroism and structural data demonstrated that the mutant isoforms did not undergo significant structural changes. Immunoassays showed that mutant PLDs exhibit conserved epitopes and kept their antigenic properties despite the mutations. Both in vitro (sphingomyelinase activity and hemolysis) and in vivo (capillary permeability, dermonecrotic activity, and histopathological analysis) assays showed that the PLDs with mutations H12-H47, E32-D34, and Y228-Y229-W230 displayed only residual activities. Results indicate that these mutant toxins are suitable for use as antigens to obtain neutralizing antisera with enhanced properties since they will be based on the most deleterious toxins in the venom and without causing severe harmful effects to the animals in which these sera are produced. |
id |
UNSP_417a7f46833b93d47c24b62d07b0c1f4 |
---|---|
oai_identifier_str |
oai:repositorio.unesp.br:11449/208589 |
network_acronym_str |
UNSP |
network_name_str |
Repositório Institucional da UNESP |
repository_id_str |
2946 |
spelling |
Brown spiders’ phospholipases-d with potential therapeutic applications: Functional assessment of mutant isoformsLoxoscelesMutant phospholipases-DPhospholipases-DSite-directed mutationsVenom enzymesPhospholipases-D (PLDs) found in Loxosceles spiders’ venoms are responsible for the der-monecrosis triggered by envenomation. PLDs can also induce other local and systemic effects, such as massive inflammatory response, edema, and hemolysis. Recombinant PLDs reproduce all of the deleterious effects induced by Loxosceles whole venoms. Herein, wild type and mutant PLDs of two species involved in accidents—L. gaucho and L. laeta—were recombinantly expressed and character-ized. The mutations are related to amino acid residues relevant for catalysis (H12-H47), magnesium ion coordination (E32-D34) and binding to phospholipid substrates (Y228 and Y228-Y229-W230). Circular dichroism and structural data demonstrated that the mutant isoforms did not undergo significant structural changes. Immunoassays showed that mutant PLDs exhibit conserved epitopes and kept their antigenic properties despite the mutations. Both in vitro (sphingomyelinase activity and hemolysis) and in vivo (capillary permeability, dermonecrotic activity, and histopathological analysis) assays showed that the PLDs with mutations H12-H47, E32-D34, and Y228-Y229-W230 displayed only residual activities. Results indicate that these mutant toxins are suitable for use as antigens to obtain neutralizing antisera with enhanced properties since they will be based on the most deleterious toxins in the venom and without causing severe harmful effects to the animals in which these sera are produced.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Departamento de Biologia Celular Universidade Federal do ParanáCentro de Produção e Pesquisa de Imunobiológicos (CPPI)Departamento de Física Centro Multiusuário de Inovação Biomolecular Universidade Estadual Paulista (UNESP)Departamento de Biologia Estrutural Molecular e Genética Universidade Estadual de Ponta GrossaDepartamento de Física Centro Multiusuário de Inovação Biomolecular Universidade Estadual Paulista (UNESP)FAPESP: 2015/13765-0FAPESP: 2019/10475-2FAPESP: 2915/18868-2CNPq: 303868/2016-3CNPq: 307338/2014-2CNPq: 309940/2019-2CNPq: 408633/2018-2Universidade Federal do Paraná (UFPR)Centro de Produção e Pesquisa de Imunobiológicos (CPPI)Universidade Estadual Paulista (Unesp)Universidade Estadual de Ponta Grossa (UEPG)da Silva, Thaís Pereirade Castro, Fernando JacominiVuitika, LarissaPolli, Nayanne Louise CostacurtaAntunes, Bruno CésarBóia-Ferreira, MariannaMinozzo, João CarlosMariutti, Ricardo Barros [UNESP]Matsubara, Fernando HitomiArni, Raghuvir Krishnaswamy [UNESP]Wille, Ana Carolina MartinsSenff-Ribeiro, AndreaGremski, Luiza HelenaVeiga, Silvio Sanches2021-06-25T11:14:38Z2021-06-25T11:14:38Z2021-03-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://dx.doi.org/10.3390/biomedicines9030320Biomedicines, v. 9, n. 3, 2021.2227-9059http://hdl.handle.net/11449/20858910.3390/biomedicines90303202-s2.0-85104266609Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengBiomedicinesinfo:eu-repo/semantics/openAccess2021-10-23T19:02:17Zoai:repositorio.unesp.br:11449/208589Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T22:11:48.515101Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Brown spiders’ phospholipases-d with potential therapeutic applications: Functional assessment of mutant isoforms |
title |
Brown spiders’ phospholipases-d with potential therapeutic applications: Functional assessment of mutant isoforms |
spellingShingle |
Brown spiders’ phospholipases-d with potential therapeutic applications: Functional assessment of mutant isoforms da Silva, Thaís Pereira Loxosceles Mutant phospholipases-D Phospholipases-D Site-directed mutations Venom enzymes |
title_short |
Brown spiders’ phospholipases-d with potential therapeutic applications: Functional assessment of mutant isoforms |
title_full |
Brown spiders’ phospholipases-d with potential therapeutic applications: Functional assessment of mutant isoforms |
title_fullStr |
Brown spiders’ phospholipases-d with potential therapeutic applications: Functional assessment of mutant isoforms |
title_full_unstemmed |
Brown spiders’ phospholipases-d with potential therapeutic applications: Functional assessment of mutant isoforms |
title_sort |
Brown spiders’ phospholipases-d with potential therapeutic applications: Functional assessment of mutant isoforms |
author |
da Silva, Thaís Pereira |
author_facet |
da Silva, Thaís Pereira de Castro, Fernando Jacomini Vuitika, Larissa Polli, Nayanne Louise Costacurta Antunes, Bruno César Bóia-Ferreira, Marianna Minozzo, João Carlos Mariutti, Ricardo Barros [UNESP] Matsubara, Fernando Hitomi Arni, Raghuvir Krishnaswamy [UNESP] Wille, Ana Carolina Martins Senff-Ribeiro, Andrea Gremski, Luiza Helena Veiga, Silvio Sanches |
author_role |
author |
author2 |
de Castro, Fernando Jacomini Vuitika, Larissa Polli, Nayanne Louise Costacurta Antunes, Bruno César Bóia-Ferreira, Marianna Minozzo, João Carlos Mariutti, Ricardo Barros [UNESP] Matsubara, Fernando Hitomi Arni, Raghuvir Krishnaswamy [UNESP] Wille, Ana Carolina Martins Senff-Ribeiro, Andrea Gremski, Luiza Helena Veiga, Silvio Sanches |
author2_role |
author author author author author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Federal do Paraná (UFPR) Centro de Produção e Pesquisa de Imunobiológicos (CPPI) Universidade Estadual Paulista (Unesp) Universidade Estadual de Ponta Grossa (UEPG) |
dc.contributor.author.fl_str_mv |
da Silva, Thaís Pereira de Castro, Fernando Jacomini Vuitika, Larissa Polli, Nayanne Louise Costacurta Antunes, Bruno César Bóia-Ferreira, Marianna Minozzo, João Carlos Mariutti, Ricardo Barros [UNESP] Matsubara, Fernando Hitomi Arni, Raghuvir Krishnaswamy [UNESP] Wille, Ana Carolina Martins Senff-Ribeiro, Andrea Gremski, Luiza Helena Veiga, Silvio Sanches |
dc.subject.por.fl_str_mv |
Loxosceles Mutant phospholipases-D Phospholipases-D Site-directed mutations Venom enzymes |
topic |
Loxosceles Mutant phospholipases-D Phospholipases-D Site-directed mutations Venom enzymes |
description |
Phospholipases-D (PLDs) found in Loxosceles spiders’ venoms are responsible for the der-monecrosis triggered by envenomation. PLDs can also induce other local and systemic effects, such as massive inflammatory response, edema, and hemolysis. Recombinant PLDs reproduce all of the deleterious effects induced by Loxosceles whole venoms. Herein, wild type and mutant PLDs of two species involved in accidents—L. gaucho and L. laeta—were recombinantly expressed and character-ized. The mutations are related to amino acid residues relevant for catalysis (H12-H47), magnesium ion coordination (E32-D34) and binding to phospholipid substrates (Y228 and Y228-Y229-W230). Circular dichroism and structural data demonstrated that the mutant isoforms did not undergo significant structural changes. Immunoassays showed that mutant PLDs exhibit conserved epitopes and kept their antigenic properties despite the mutations. Both in vitro (sphingomyelinase activity and hemolysis) and in vivo (capillary permeability, dermonecrotic activity, and histopathological analysis) assays showed that the PLDs with mutations H12-H47, E32-D34, and Y228-Y229-W230 displayed only residual activities. Results indicate that these mutant toxins are suitable for use as antigens to obtain neutralizing antisera with enhanced properties since they will be based on the most deleterious toxins in the venom and without causing severe harmful effects to the animals in which these sera are produced. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021-06-25T11:14:38Z 2021-06-25T11:14:38Z 2021-03-01 |
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/10.3390/biomedicines9030320 Biomedicines, v. 9, n. 3, 2021. 2227-9059 http://hdl.handle.net/11449/208589 10.3390/biomedicines9030320 2-s2.0-85104266609 |
url |
http://dx.doi.org/10.3390/biomedicines9030320 http://hdl.handle.net/11449/208589 |
identifier_str_mv |
Biomedicines, v. 9, n. 3, 2021. 2227-9059 10.3390/biomedicines9030320 2-s2.0-85104266609 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Biomedicines |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.source.none.fl_str_mv |
Scopus 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_ |
1808129403516878848 |