Purification and characterization of the first gamma-phospholipase inhibitor (gamma PLI) from Bothrops jararaca snake serum

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dc.contributorLaboratório de Herpetologiapt_BR
dc.contributorCentro de Biotecnologiapt_BR
dc.contributor.authorSilva, Caroline Serinopt_BR
dc.contributor.authorMorais-Zani, Karen dept_BR
dc.contributor.authorToyama, Marcos Hikaript_BR
dc.contributor.authorToyama, Daniela de Oliveirapt_BR
dc.contributor.authorGaeta, Henrique Hessept_BR
dc.contributor.authorFabri, Carolinept_BR
dc.contributor.authorAguiar, Weslei da Silvapt_BR
dc.contributor.authorTashima, Alexandre K.pt_BR
dc.contributor.authorGrego, Kathleen Fernandespt_BR
dc.contributor.authorTanaka-Azevedo, Anita Miticopt_BR
dc.date.accessioned2020-07-09T21:19:12Z-
dc.date.available2020-07-09T21:19:12Z-
dc.date.issued2018-
dc.identifier.citationSilva CS, Morais-Zani K, Toyama MH, Toyama DO, Gaeta HH, Fabri C, et al. Purification and characterization of the first gamma-phospholipase inhibitor (gamma PLI) from Bothrops jararaca snake serum. PLoS One. 2018 Mar:13(3):e0193105. doi:10.1371/journal.pone.0193105.pt_BR
dc.identifier.issn1932-6203-
dc.identifier.urihttps://repositorio.butantan.gov.br/handle/butantan/2404-
dc.description.abstractPhospholipases A(2) (PLA(2)) are enzymes acting on the cell membrane phospholipids resulting in fatty acids and lysophospholipids and deconstructing the cell membrane. This protein is commonly found in snake venoms, causing tissue inflammation in the affected area. Evidence indicates that snakes have natural resistance to their own venom due to protective properties in plasma, that inhibit the action of proteins present in their venom. Given that, this study aimed to purify and characterize a gamma PLI from Bothrops jararaca serum, named gamma BjPLI. PLA(2) inhibitor was isolated using two chromatographic steps: an ion exchange column (DEAE), followed by an affinity column (crotoxin coupled to a CNBr-activated Sepharose resin). The purity and biochemical characterization of the isolated protein were analyzed by RP-HPLC, SEC, SDS-PAGE, circular dichroism and mass spectrometry. The ability to inhibit PLA(2) was determined by enzymatic activity, neutralization of paw edema and myonecrosis. The protein purity was confirmed by RP-HPLC and SEC, whilst an apparent molecular mass of 25 kDa and 20 kDa was obtained by SDS-PAGE, under reducing and non-reducing conditions, respectively. According to mass spectrometry analysis, this protein showed 72% and 68% of coverage when aligned to amino acid sequences of two proteins already described as PLIs. Thus, the inhibitory activity of enzymatic, edema and myonecrotic activities by gamma BjPLI suggests a role of this inhibitor for protection of these snakes against self-envenomation.pt_BR
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)pt_BR
dc.description.sponsorshipCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)pt_BR
dc.format.extente0193105pt_BR
dc.languageengpt_BR
dc.publisherPublic Library Sciencept_BR
dc.relation.ispartofPloS Onept_BR
dc.rightsOpen Accesspt_BR
dc.titlePurification and characterization of the first gamma-phospholipase inhibitor (gamma PLI) from Bothrops jararaca snake serumpt_BR
dc.typeArticlept_BR
dc.identifier.doi10.1371/journal.pone.0193105pt_BR
dc.identifier.urlhttps://dx.doi.org/10.1371/journal.pone.0193105pt_BR
dc.contributor.externalUniversidade de São Paulo (USP)¦¦Brasilpt_BR
dc.contributor.externalUniversidade Federal de São Paulo (UNIFESP)¦¦Brasilpt_BR
dc.contributor.externalUniversidade Estadual Paulista Júlio de Mesquita Filho (UNESP)¦¦Brasilpt_BR
dc.publisher.citySan Franciscopt_BR
dc.identifier.citationvolume13pt_BR
dc.identifier.citationissue3pt_BR
dc.relation.ispartofabbreviatedPLoS Onept_BR
dc.identifier.citationabntv. 13, n. 3, e0193105, mar. 2018pt_BR
dc.identifier.citationvancouver2018 Mar:13(3):e0193105pt_BR
dc.publisher.countryUnited Statespt_BR
dc.contributor.butantanMorais-Zani, Karen de|:Pesquisador|:Laboratório de Herpetologia|:pt_BR
dc.contributor.butantanFabri, Caroline|:Aluno|:Laboratório de Herpetologia|:pt_BR
dc.contributor.butantanAguiar, Weslei da Silva|:Aluno|:Centro de Biotecnologia|:pt_BR
dc.contributor.butantanGrego, Kathleen Fernandes|:Pesquisador|:Laboratório de Herpetologia|:pt_BR
dc.contributor.butantanTanaka-Azevedo, Anita Mitico|:Pesquisador|:Laboratório de Herpetologia|:Autor de correspondênciapt_BR
dc.contributor.butantanSilva, Caroline Serino|:Aluno|:Laboratório de Herpetologia|:PrimeiroAutorpt_BR
dc.sponsorship.butantanCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)¦¦pt_BR
dc.sponsorship.butantanFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)¦¦2012/19321-9pt_BR
dc.sponsorship.butantanFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)¦¦2016/03839-0pt_BR
dc.sponsorship.butantanFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)¦¦2013/05357-4pt_BR
dc.sponsorship.butantanFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)¦¦2014/11108-0pt_BR
dc.sponsorship.butantanFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)¦¦2017/01890-0pt_BR
dc.sponsorship.butantanFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)¦¦2017/16908-2pt_BR
dc.sponsorship.butantanFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)¦¦2013/12826-0pt_BR
dc.identifier.bvsccBR78.1pt_BR
dc.identifier.bvsdbIBProdpt_BR
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