Mechanistic insights into functional characteristics of native crotamine
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Outros Autores: | , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1016/j.toxicon.2018.03.007 http://hdl.handle.net/11449/170828 |
Resumo: | The chemical composition of snake venoms is a complex mixture of proteins and peptides that can be pharmacologically active. Crotamine, a cell-penetrating peptide, has been described to have antimicrobial properties and it exerts its effects by interacting selectively with different structures, inducing changes in the ion flow pattern and cellular responses. However, its real therapeutic potential is not yet fully known. Bearing in mind that crotamine is a promising molecule in therapeutics, this study investigated the action of purified molecule in three aspects: I) antibacterial action on different species of clinical interest, II) the effect of two different concentrations of the molecule on platelet aggregation, and III) its effects on isolated mitochondria. Crotamine was purified to homogeneity in a single step procedure using Heparin Sepharose. The molecular mass of the purified enzyme was 4881.4 Da, as determined by mass spectrometry. To assess antibacterial action, changes in the parameters of bacterial oxidative stress were determined. The peptide showed antibacterial activity on Escherichia coli (MIC: 2.0 μg/μL), Staphylococcus aureus (MIC: 8–16 μg/μL) and methicillin-resistant Staphylococcus aureus (MIC: 4.0–8.0 μg/μL), inducing bacterial death by lipid peroxidation and oxidation of target proteins, determined by thiobarbituric acid reactive substances and sulfhydryl groups, respectively. Crotamine induced increased platelet aggregation (IPA) at the two concentrations analyzed (0.1 and 1.4 μg/μL) compared to ADP-induced aggregation of PRP. Mitochondrial respiratory parameters and organelle structure assays were used to elucidate the action of the compound in this organelle. The exposure of mitochondria to crotamine caused a decrease in oxidative phosphorylation and changes in mitochondrial permeability, without causing damage in the mitochondrial redox state. Together, these results support the hypothesis that, besides the antimicrobial potential, crotamine acts on different molecular targets, inducing platelet aggregation and mitochondrial dysfunction. |
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Mechanistic insights into functional characteristics of native crotamineAntimicrobialCrotalus durissus terrificusCrotamineMitochondrial dysfunctionPlatelet aggregationThe chemical composition of snake venoms is a complex mixture of proteins and peptides that can be pharmacologically active. Crotamine, a cell-penetrating peptide, has been described to have antimicrobial properties and it exerts its effects by interacting selectively with different structures, inducing changes in the ion flow pattern and cellular responses. However, its real therapeutic potential is not yet fully known. Bearing in mind that crotamine is a promising molecule in therapeutics, this study investigated the action of purified molecule in three aspects: I) antibacterial action on different species of clinical interest, II) the effect of two different concentrations of the molecule on platelet aggregation, and III) its effects on isolated mitochondria. Crotamine was purified to homogeneity in a single step procedure using Heparin Sepharose. The molecular mass of the purified enzyme was 4881.4 Da, as determined by mass spectrometry. To assess antibacterial action, changes in the parameters of bacterial oxidative stress were determined. The peptide showed antibacterial activity on Escherichia coli (MIC: 2.0 μg/μL), Staphylococcus aureus (MIC: 8–16 μg/μL) and methicillin-resistant Staphylococcus aureus (MIC: 4.0–8.0 μg/μL), inducing bacterial death by lipid peroxidation and oxidation of target proteins, determined by thiobarbituric acid reactive substances and sulfhydryl groups, respectively. Crotamine induced increased platelet aggregation (IPA) at the two concentrations analyzed (0.1 and 1.4 μg/μL) compared to ADP-induced aggregation of PRP. Mitochondrial respiratory parameters and organelle structure assays were used to elucidate the action of the compound in this organelle. The exposure of mitochondria to crotamine caused a decrease in oxidative phosphorylation and changes in mitochondrial permeability, without causing damage in the mitochondrial redox state. Together, these results support the hypothesis that, besides the antimicrobial potential, crotamine acts on different molecular targets, inducing platelet aggregation and mitochondrial dysfunction.Graduate Program in Biomedical Sciences Hermínio Ometto University Center UNIARARAS, 7 Av. Dr. Maximiliano Baruto, 500Graduate Program in Agrarian and Veterinary Sciences State University Paulista Júlio de Mesquita Filho-UNESPBlood Hemostasis Laboratory Faculty of Medical Sciences State University of CampinasDepartment of Bioprocesses and Biotechnology Faculty of Agronomic Sciences State University Paulista Júlio Mesquita Filho-UNESPGraduate Program in Agrarian and Veterinary Sciences State University Paulista Júlio de Mesquita Filho-UNESPDepartment of Bioprocesses and Biotechnology Faculty of Agronomic Sciences State University Paulista Júlio Mesquita Filho-UNESPUNIARARASUniversidade Estadual Paulista (Unesp)Universidade Estadual de Campinas (UNICAMP)Batista da Cunha, DanielPupo Silvestrini, Ana VitóriaGomes da Silva, Ana CarolinaMaria de Paula Estevam, Deborah [UNESP]Pollettini, Flávia Lino [UNESP]de Oliveira Navarro, JulianaAlves, Armindo AntônioRemédio Zeni Beretta, Ana LauraAnnichino Bizzacchi, Joyce M.Pereira, Lilian Cristina [UNESP]Mazzi, Maurício Ventura2018-12-11T16:52:35Z2018-12-11T16:52:35Z2018-05-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article1-12application/pdfhttp://dx.doi.org/10.1016/j.toxicon.2018.03.007Toxicon, v. 146, p. 1-12.1879-31500041-0101http://hdl.handle.net/11449/17082810.1016/j.toxicon.2018.03.0072-s2.0-850444687332-s2.0-85044468733.pdfScopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengToxicon0,692info:eu-repo/semantics/openAccess2024-01-12T06:27:19Zoai:repositorio.unesp.br:11449/170828Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-01-12T06:27:19Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Mechanistic insights into functional characteristics of native crotamine |
title |
Mechanistic insights into functional characteristics of native crotamine |
spellingShingle |
Mechanistic insights into functional characteristics of native crotamine Batista da Cunha, Daniel Antimicrobial Crotalus durissus terrificus Crotamine Mitochondrial dysfunction Platelet aggregation |
title_short |
Mechanistic insights into functional characteristics of native crotamine |
title_full |
Mechanistic insights into functional characteristics of native crotamine |
title_fullStr |
Mechanistic insights into functional characteristics of native crotamine |
title_full_unstemmed |
Mechanistic insights into functional characteristics of native crotamine |
title_sort |
Mechanistic insights into functional characteristics of native crotamine |
author |
Batista da Cunha, Daniel |
author_facet |
Batista da Cunha, Daniel Pupo Silvestrini, Ana Vitória Gomes da Silva, Ana Carolina Maria de Paula Estevam, Deborah [UNESP] Pollettini, Flávia Lino [UNESP] de Oliveira Navarro, Juliana Alves, Armindo Antônio Remédio Zeni Beretta, Ana Laura Annichino Bizzacchi, Joyce M. Pereira, Lilian Cristina [UNESP] Mazzi, Maurício Ventura |
author_role |
author |
author2 |
Pupo Silvestrini, Ana Vitória Gomes da Silva, Ana Carolina Maria de Paula Estevam, Deborah [UNESP] Pollettini, Flávia Lino [UNESP] de Oliveira Navarro, Juliana Alves, Armindo Antônio Remédio Zeni Beretta, Ana Laura Annichino Bizzacchi, Joyce M. Pereira, Lilian Cristina [UNESP] Mazzi, Maurício Ventura |
author2_role |
author author author author author author author author author author |
dc.contributor.none.fl_str_mv |
UNIARARAS Universidade Estadual Paulista (Unesp) Universidade Estadual de Campinas (UNICAMP) |
dc.contributor.author.fl_str_mv |
Batista da Cunha, Daniel Pupo Silvestrini, Ana Vitória Gomes da Silva, Ana Carolina Maria de Paula Estevam, Deborah [UNESP] Pollettini, Flávia Lino [UNESP] de Oliveira Navarro, Juliana Alves, Armindo Antônio Remédio Zeni Beretta, Ana Laura Annichino Bizzacchi, Joyce M. Pereira, Lilian Cristina [UNESP] Mazzi, Maurício Ventura |
dc.subject.por.fl_str_mv |
Antimicrobial Crotalus durissus terrificus Crotamine Mitochondrial dysfunction Platelet aggregation |
topic |
Antimicrobial Crotalus durissus terrificus Crotamine Mitochondrial dysfunction Platelet aggregation |
description |
The chemical composition of snake venoms is a complex mixture of proteins and peptides that can be pharmacologically active. Crotamine, a cell-penetrating peptide, has been described to have antimicrobial properties and it exerts its effects by interacting selectively with different structures, inducing changes in the ion flow pattern and cellular responses. However, its real therapeutic potential is not yet fully known. Bearing in mind that crotamine is a promising molecule in therapeutics, this study investigated the action of purified molecule in three aspects: I) antibacterial action on different species of clinical interest, II) the effect of two different concentrations of the molecule on platelet aggregation, and III) its effects on isolated mitochondria. Crotamine was purified to homogeneity in a single step procedure using Heparin Sepharose. The molecular mass of the purified enzyme was 4881.4 Da, as determined by mass spectrometry. To assess antibacterial action, changes in the parameters of bacterial oxidative stress were determined. The peptide showed antibacterial activity on Escherichia coli (MIC: 2.0 μg/μL), Staphylococcus aureus (MIC: 8–16 μg/μL) and methicillin-resistant Staphylococcus aureus (MIC: 4.0–8.0 μg/μL), inducing bacterial death by lipid peroxidation and oxidation of target proteins, determined by thiobarbituric acid reactive substances and sulfhydryl groups, respectively. Crotamine induced increased platelet aggregation (IPA) at the two concentrations analyzed (0.1 and 1.4 μg/μL) compared to ADP-induced aggregation of PRP. Mitochondrial respiratory parameters and organelle structure assays were used to elucidate the action of the compound in this organelle. The exposure of mitochondria to crotamine caused a decrease in oxidative phosphorylation and changes in mitochondrial permeability, without causing damage in the mitochondrial redox state. Together, these results support the hypothesis that, besides the antimicrobial potential, crotamine acts on different molecular targets, inducing platelet aggregation and mitochondrial dysfunction. |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-12-11T16:52:35Z 2018-12-11T16:52:35Z 2018-05-01 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1016/j.toxicon.2018.03.007 Toxicon, v. 146, p. 1-12. 1879-3150 0041-0101 http://hdl.handle.net/11449/170828 10.1016/j.toxicon.2018.03.007 2-s2.0-85044468733 2-s2.0-85044468733.pdf |
url |
http://dx.doi.org/10.1016/j.toxicon.2018.03.007 http://hdl.handle.net/11449/170828 |
identifier_str_mv |
Toxicon, v. 146, p. 1-12. 1879-3150 0041-0101 10.1016/j.toxicon.2018.03.007 2-s2.0-85044468733 2-s2.0-85044468733.pdf |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Toxicon 0,692 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
1-12 application/pdf |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
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UNESP |
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UNESP |
reponame_str |
Repositório Institucional da UNESP |
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Repositório Institucional da UNESP |
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Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
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