Brown spiders’ phospholipases-d with potential therapeutic applications: Functional assessment of mutant isoforms

Detalhes bibliográficos
Autor(a) principal: da Silva, Thaís Pereira
Data de Publicação: 2021
Outros Autores: 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
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.
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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
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