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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...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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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. |
format | Online Article Text |
id | pubmed-8004160 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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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