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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

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

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Author(s):
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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]
Total Authors: 14
Affiliation:
[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
Total Affiliations: 4
Document type: Journal article
Source: BIOMEDICINES; v. 9, n. 3 MAR 2021.
Web of Science Citations: 0
Abstract

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)

FAPESP's process: 15/18868-2 - Multi-user equipment acquisition for molecular and structural biology
Grantee:Raghuvir Krishnaswamy Arni
Support Opportunities: Multi-user Equipment Program
FAPESP's process: 15/13765-0 - Structural Studies and Characterization of Proteins by X-ray Crystallography and Nuclear Magnetic Resonance. Structural investigations and biophysics of molecular mechanisms of functional proteins.
Grantee:Raghuvir Krishnaswamy Arni
Support Opportunities: Regular Research Grants