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

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 i...

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Autores principales: 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, Matsubara, Fernando Hitomi, Arni, Raghuvir Krishnaswamy, Wille, Ana Carolina Martins, Senff-Ribeiro, Andrea, Gremski, Luiza Helena, Veiga, Silvio Sanches
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8004160/
https://www.ncbi.nlm.nih.gov/pubmed/33801128
http://dx.doi.org/10.3390/biomedicines9030320
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author 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
Matsubara, Fernando Hitomi
Arni, Raghuvir Krishnaswamy
Wille, Ana Carolina Martins
Senff-Ribeiro, Andrea
Gremski, Luiza Helena
Veiga, Silvio Sanches
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
Matsubara, Fernando Hitomi
Arni, Raghuvir Krishnaswamy
Wille, Ana Carolina Martins
Senff-Ribeiro, Andrea
Gremski, Luiza Helena
Veiga, Silvio Sanches
author_sort da Silva, Thaís Pereira
collection PubMed
description 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.
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spelling pubmed-80041602021-03-28 Brown Spiders’ Phospholipases-D with Potential Therapeutic Applications: Functional Assessment of Mutant Isoforms 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 Matsubara, Fernando Hitomi Arni, Raghuvir Krishnaswamy Wille, Ana Carolina Martins Senff-Ribeiro, Andrea Gremski, Luiza Helena Veiga, Silvio Sanches Biomedicines Article 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. MDPI 2021-03-21 /pmc/articles/PMC8004160/ /pubmed/33801128 http://dx.doi.org/10.3390/biomedicines9030320 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ).
spellingShingle Article
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
Matsubara, Fernando Hitomi
Arni, Raghuvir Krishnaswamy
Wille, Ana Carolina Martins
Senff-Ribeiro, Andrea
Gremski, Luiza Helena
Veiga, Silvio Sanches
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
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_short 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
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8004160/
https://www.ncbi.nlm.nih.gov/pubmed/33801128
http://dx.doi.org/10.3390/biomedicines9030320
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