Brown spiders’ phospholipases-d with potential therapeutic applications: Functional assessment of mutant isoforms
dc.contributor.author | da Silva, Thaís Pereira | |
dc.contributor.author | de Castro, Fernando Jacomini | |
dc.contributor.author | Vuitika, Larissa | |
dc.contributor.author | Polli, Nayanne Louise Costacurta | |
dc.contributor.author | Antunes, Bruno César | |
dc.contributor.author | Bóia-Ferreira, Marianna | |
dc.contributor.author | Minozzo, João Carlos | |
dc.contributor.author | Mariutti, Ricardo Barros [UNESP] | |
dc.contributor.author | Matsubara, Fernando Hitomi | |
dc.contributor.author | Arni, Raghuvir Krishnaswamy [UNESP] | |
dc.contributor.author | Wille, Ana Carolina Martins | |
dc.contributor.author | Senff-Ribeiro, Andrea | |
dc.contributor.author | Gremski, Luiza Helena | |
dc.contributor.author | Veiga, Silvio Sanches | |
dc.contributor.institution | Universidade Federal do Paraná (UFPR) | |
dc.contributor.institution | Centro de Produção e Pesquisa de Imunobiológicos (CPPI) | |
dc.contributor.institution | Universidade Estadual Paulista (Unesp) | |
dc.contributor.institution | Universidade Estadual de Ponta Grossa (UEPG) | |
dc.date.accessioned | 2021-06-25T11:14:38Z | |
dc.date.available | 2021-06-25T11:14:38Z | |
dc.date.issued | 2021-03-01 | |
dc.description.abstract | 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. | en |
dc.description.affiliation | Departamento de Biologia Celular Universidade Federal do Paraná | |
dc.description.affiliation | Centro de Produção e Pesquisa de Imunobiológicos (CPPI) | |
dc.description.affiliation | Departamento de Física Centro Multiusuário de Inovação Biomolecular Universidade Estadual Paulista (UNESP) | |
dc.description.affiliation | Departamento de Biologia Estrutural Molecular e Genética Universidade Estadual de Ponta Grossa | |
dc.description.affiliationUnesp | Departamento de Física Centro Multiusuário de Inovação Biomolecular Universidade Estadual Paulista (UNESP) | |
dc.description.sponsorship | Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) | |
dc.description.sponsorship | Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) | |
dc.description.sponsorshipId | FAPESP: 2015/13765-0 | |
dc.description.sponsorshipId | FAPESP: 2019/10475-2 | |
dc.description.sponsorshipId | FAPESP: 2915/18868-2 | |
dc.description.sponsorshipId | CNPq: 303868/2016-3 | |
dc.description.sponsorshipId | CNPq: 307338/2014-2 | |
dc.description.sponsorshipId | CNPq: 309940/2019-2 | |
dc.description.sponsorshipId | CNPq: 408633/2018-2 | |
dc.identifier | http://dx.doi.org/10.3390/biomedicines9030320 | |
dc.identifier.citation | Biomedicines, v. 9, n. 3, 2021. | |
dc.identifier.doi | 10.3390/biomedicines9030320 | |
dc.identifier.issn | 2227-9059 | |
dc.identifier.scopus | 2-s2.0-85104266609 | |
dc.identifier.uri | http://hdl.handle.net/11449/208589 | |
dc.language.iso | eng | |
dc.relation.ispartof | Biomedicines | |
dc.source | Scopus | |
dc.subject | Loxosceles | |
dc.subject | Mutant phospholipases-D | |
dc.subject | Phospholipases-D | |
dc.subject | Site-directed mutations | |
dc.subject | Venom enzymes | |
dc.title | Brown spiders’ phospholipases-d with potential therapeutic applications: Functional assessment of mutant isoforms | en |
dc.type | Artigo | |
dspace.entity.type | Publication | |
unesp.campus | Universidade Estadual Paulista (UNESP), Instituto de Biociências, Letras e Ciências Exatas, São José do Rio Preto | pt |
unesp.department | Física - IBILCE | pt |