Harpalycin 2 inhibits the enzymatic and platelet aggregation activities of PrTX-III, a D49 phospholipase A(2) from Bothrops pirajai venom

Página do item simplificado
dc.contributor.authorXimenes, Rafael M.
dc.contributor.authorAlves, Renata S.
dc.contributor.authorPereira, Ticiana P.
dc.contributor.authorAraujo, Renata M.
dc.contributor.authorSilveira, Edilberto R.
dc.contributor.authorRabello, Marcelo M.
dc.contributor.authorHernandes, Marcelo Z.
dc.contributor.authorSoares, Veronica C. G. [UNESP]
dc.contributor.authorBristot, Daniel [UNESP]
dc.contributor.authorPires, Camila L. [UNESP]
dc.contributor.authorToyama, Daniela O.
dc.contributor.authorGaeta, Henrique H. [UNESP]
dc.contributor.authorMonteiro, Helena S. A.
dc.contributor.authorToyama, Marcos H. [UNESP]
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.contributor.institutionUniversidade Federal do Ceará (UFC)
dc.contributor.institutionUniversidade Federal do Rio Grande do Norte (UFRN)
dc.contributor.institutionUniversidade Federal de Pernambuco (UFPE)
dc.contributor.institutionUniversidade Estadual de Campinas (UNICAMP)
dc.contributor.institutionUniv Prebiteriana Mackenzie
dc.date.accessioned2014-05-20T13:12:24Z
dc.date.available2014-05-20T13:12:24Z
dc.date.issued2012-08-27
dc.description.abstractBackground: Harpalycin 2 (HP-2) is an isoflavone isolated from the leaves of Harpalyce brasiliana Benth., a snakeroot found in northeast region of Brazil and used in folk medicine to treat snakebite. Its leaves are said to be anti-inflammatory. Secretory phospholipases A(2) are important toxins found in snake venom and are structurally related to those found in inflammatory conditions in mammals, as in arthritis and atherosclerosis, and for this reason can be valuable tools for searching new anti-phospholipase A(2) drugs.Methods: HP-2 and piratoxin-III (PrTX-III) were purified through chromatographic techniques. The effect of HP-2 in the enzymatic activity of PrTX-III was carried out using 4-nitro-3-octanoyloxy-benzoic acid as the substrate. PrTX-III induced platelet aggregation was inhibited by HP-2 when compared to aristolochic acid and p-bromophenacyl bromide (p-BPB). In an attempt to elucidate how HP-2 interacts with PrTX-III, mass spectrometry, circular dichroism and intrinsic fluorescence analysis were performed. Docking scores of the ligands (HP-2, aristolochic acid and p-BPB) using PrTX-III as target were also calculated.Results: HP-2 inhibited the enzymatic activity of PrTX-III (IC50 11.34 +/- 0.28 mu g/mL) although it did not form a stable chemical complex in the active site, since mass spectrometry measurements showed no difference between native (13,837.34 Da) and HP-2 treated PrTX-III (13,856.12 Da). A structural analysis of PrTX-III after treatment with HP-2 showed a decrease in dimerization and a slight protein unfolding. In the platelet aggregation assay, HP-2 previously incubated with PrTX-III inhibited the aggregation when compared with untreated protein. PrTX-III chemical treated with aristolochic acid and p-BPB, two standard PLA(2) inhibitors, showed low inhibitory effects when compared with the HP-2 treatment. Docking scores corroborated these results, showing higher affinity of HP-2 for the PrTX-III target (PDB code: 1GMZ) than aristolochic acid and p-BPB. HP-2 previous incubated with the platelets inhibits the aggregation induced by untreated PrTX-III as well as arachidonic acid.Conclusion: HP-2 changes the structure of PrTX-III, inhibiting the enzymatic activity of this enzyme. In addition, PrTX-III platelet aggregant activity was inhibited by treatment with HP-2, p-BPB and aristolochic acid, and these results were corroborated by docking scores.en
dc.description.affiliationState Univ São Paulo Julio Mesquita Filho, UNESP, Sao Vicente Unit, BR-11330900 Sao Vicente, SP, Brazil
dc.description.affiliationUniv Fed Ceara, UFC, Dept Physiol & Pharmacol, BR-60430270 Fortaleza, Ceara, Brazil
dc.description.affiliationUniv Fed Ceara, UFC, Dept Clin & Toxicol Anal, BR-60430370 Fortaleza, Ceara, Brazil
dc.description.affiliationUniv Fed Rio Grande do Norte, UFRN, Dept Chem, BR-50078970 Natal, RN, Brazil
dc.description.affiliationUniv Fed Ceara, Dept Organ & Inorgan Chem, UFC, BR-60455760 Fortaleza, Ceara, Brazil
dc.description.affiliationUniversidade Federal de Pernambuco (UFPE), UFPE, Dept Pharmaceut Sci, BR-50740520 Recife, PE, Brazil
dc.description.affiliationUniv Estadual Campinas, UNICAMP, Inst Biol, Dept Biochem, BR-13082862 Campinas, SP, Brazil
dc.description.affiliationUniv Prebiteriana Mackenzie, Ctr Biol & Hlth Sci, BR-01302970 São Paulo, Brazil
dc.description.affiliationUnespState Univ São Paulo Julio Mesquita Filho, UNESP, Sao Vicente Unit, BR-11330900 Sao Vicente, SP, Brazil
dc.description.sponsorshipCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipFundação Cearense de Apoio ao Desenvolvimento Científico e Tecnológico (FUNCAP)
dc.format.extent10
dc.identifierhttp://dx.doi.org/10.1186/1472-6882-12-139
dc.identifier.citationBmc Complementary and Alternative Medicine. London: Biomed Central Ltd., v. 12, p. 10, 2012.
dc.identifier.doi10.1186/1472-6882-12-139
dc.identifier.fileWOS000312325000001.pdf
dc.identifier.issn1472-6882
dc.identifier.urihttp://hdl.handle.net/11449/382
dc.identifier.wosWOS:000312325000001
dc.language.isoeng
dc.publisherBiomed Central Ltd.
dc.relation.ispartofBMC Complementary and Alternative Medicine
dc.relation.ispartofjcr2.109
dc.relation.ispartofsjr0,858
dc.rights.accessRightsAcesso aberto
dc.sourceWeb of Science
dc.subjectPrTX-IIIen
dc.subjectPhospholipase A(2)en
dc.subjectBothrops pirajaien
dc.subjectHarpalyce brasilianaen
dc.subjectIsoflavoneen
dc.titleHarpalycin 2 inhibits the enzymatic and platelet aggregation activities of PrTX-III, a D49 phospholipase A(2) from Bothrops pirajai venomen
dc.typeArtigo
dcterms.licensehttp://www.biomedcentral.com/about/license
dcterms.rightsHolderBiomed Central Ltd.
unesp.campusUniversidade Estadual Paulista (Unesp), Instituto de Biociências, São Vicentept

Arquivos

Pacote Original
Agora exibindo 1 - 1 de 1
Imagem de Miniatura
Nome:
WOS000312325000001.pdf
Tamanho:
1.61 MB
Formato:
Adobe Portable Document Format
Baixar
Licença do Pacote
Agora exibindo 1 - 2 de 2
Nenhuma Miniatura disponível
Nome:
license.txt
Tamanho:
1.71 KB
Formato:
Item-specific license agreed upon to submission
Descrição:
Nenhuma Miniatura disponível
Nome:
license.txt
Tamanho:
1.71 KB
Formato:
Item-specific license agreed upon to submission
Descrição:

Coleções

Artigos - Ciências Biológicas - São Vicente