Logo do repositório
 

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

dc.contributor.authorda Silva, Thaís Pereira
dc.contributor.authorde Castro, Fernando Jacomini
dc.contributor.authorVuitika, Larissa
dc.contributor.authorPolli, Nayanne Louise Costacurta
dc.contributor.authorAntunes, Bruno César
dc.contributor.authorBóia-Ferreira, Marianna
dc.contributor.authorMinozzo, João Carlos
dc.contributor.authorMariutti, Ricardo Barros [UNESP]
dc.contributor.authorMatsubara, Fernando Hitomi
dc.contributor.authorArni, Raghuvir Krishnaswamy [UNESP]
dc.contributor.authorWille, Ana Carolina Martins
dc.contributor.authorSenff-Ribeiro, Andrea
dc.contributor.authorGremski, Luiza Helena
dc.contributor.authorVeiga, Silvio Sanches
dc.contributor.institutionUniversidade Federal do Paraná (UFPR)
dc.contributor.institutionCentro de Produção e Pesquisa de Imunobiológicos (CPPI)
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.contributor.institutionUniversidade Estadual de Ponta Grossa (UEPG)
dc.date.accessioned2021-06-25T11:14:38Z
dc.date.available2021-06-25T11:14:38Z
dc.date.issued2021-03-01
dc.description.abstractPhospholipases-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.en
dc.description.affiliationDepartamento de Biologia Celular Universidade Federal do Paraná
dc.description.affiliationCentro de Produção e Pesquisa de Imunobiológicos (CPPI)
dc.description.affiliationDepartamento de Física Centro Multiusuário de Inovação Biomolecular Universidade Estadual Paulista (UNESP)
dc.description.affiliationDepartamento de Biologia Estrutural Molecular e Genética Universidade Estadual de Ponta Grossa
dc.description.affiliationUnespDepartamento de Física Centro Multiusuário de Inovação Biomolecular Universidade Estadual Paulista (UNESP)
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.sponsorshipIdFAPESP: 2015/13765-0
dc.description.sponsorshipIdFAPESP: 2019/10475-2
dc.description.sponsorshipIdFAPESP: 2915/18868-2
dc.description.sponsorshipIdCNPq: 303868/2016-3
dc.description.sponsorshipIdCNPq: 307338/2014-2
dc.description.sponsorshipIdCNPq: 309940/2019-2
dc.description.sponsorshipIdCNPq: 408633/2018-2
dc.identifierhttp://dx.doi.org/10.3390/biomedicines9030320
dc.identifier.citationBiomedicines, v. 9, n. 3, 2021.
dc.identifier.doi10.3390/biomedicines9030320
dc.identifier.issn2227-9059
dc.identifier.scopus2-s2.0-85104266609
dc.identifier.urihttp://hdl.handle.net/11449/208589
dc.language.isoeng
dc.relation.ispartofBiomedicines
dc.sourceScopus
dc.subjectLoxosceles
dc.subjectMutant phospholipases-D
dc.subjectPhospholipases-D
dc.subjectSite-directed mutations
dc.subjectVenom enzymes
dc.titleBrown spiders’ phospholipases-d with potential therapeutic applications: Functional assessment of mutant isoformsen
dc.typeArtigo
dspace.entity.typePublication
unesp.campusUniversidade Estadual Paulista (UNESP), Instituto de Biociências, Letras e Ciências Exatas, São José do Rio Pretopt
unesp.departmentFísica - IBILCEpt

Arquivos