Brown Spiders' Phospholipases-D with Potential Therapeutic Applications: Functional Assessment of Mutant Isoforms

Autor(es):Mostrar menos -

da Silva, Thais Pereira ^[1] ; de Castro, Fernando Jacomini ^[1] ; Vuitika, Larissa ^[1] ; Polli, Nayanne Louise Costacurta ^[1] ; Antunes, Bruno Cesar ^{[1, 2]} ; Boia-Ferreira, Marianna ^[1] ; Minozzo, Joao Carlos ^[2] ; Mariutti, Ricardo Barros ^[3] ; Matsubara, Fernando Hitomi ^[1] ; Arni, Raghuvir Krishnaswamy ^[3] ; Wille, Ana Carolina Martins ^[4] ; Senff-Ribeiro, Andrea ^[1] ; Gremski, Luiza Helena ^[1] ; Veiga, Silvio Sanches ^[1]

Afiliação do(s) autor(es)

^[1] Univ Fed Parana, Dept Biol Celular, BR-81530900 Curitiba, Parana - Brazil

^[2] Ctr Prod & Pesquisa Imunobiol CPPI, BR-83302200 Piraquara, Parana - Brazil

^[3] Univ Estadual Paulista UNESP, Ctr Multiusuario Inovacao Biomol, Dept Fis, BR-15054000 Sao Jose Do Rio Preto, SP - Brazil

^[4] Univ Estadual Ponta Grossa, Dept Biol Estrutural Mol & Genet, BR-84030900 Ponta Grossa, Parana - Brazil

• Tipo de documento: Artigo Científico
• Fonte: BIOMEDICINES; v. 9, n. 3 MAR 2021.
• Citações Web of Science: 0

Resumo

Phospholipases-D (PLDs) found in Loxosceles spiders' venoms are responsible for the dermonecrosis 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 characterized. 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. (AU)

• Processo FAPESP: 15/18868-2 - Aquisição de equipamento multiusuário para biologia molecular e estrutural
• Beneficiário: Raghuvir Krishnaswamy Arni
• Modalidade de apoio: Auxílio à Pesquisa - Programa Equipamentos Multiusuários
• Processo FAPESP: 15/13765-0 - ESTUDOS ESTRUTURAIS E CARACTERIZAÇÃO DE PROTEÍNAS POR CRISTALOGRAFIA DE RAIOS-X E RESSONÂNCIA MAGNETICA NUCLEAR. Investigações estruturais e biofísicas dos mecanismos moleculares de proteínas funcionais
• Beneficiário: Raghuvir Krishnaswamy Arni
• Modalidade de apoio: Auxílio à Pesquisa - Regular