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

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dc.citation.issue3
dc.citation.volumev. 13
dc.contributor.authorSerino-Silva, Caroline
dc.contributor.authorMorais-Zani, Karen
dc.contributor.authorToyama, Marcos Hikari
dc.contributor.authorToyama, Daniela de Oliveira
dc.contributor.authorGaeta, Henrique Hesse
dc.contributor.authorRodrigues, Caroline Fabri Bittencourt
dc.contributor.authorAguiar, Weslei da Silva
dc.contributor.authorTashima, Alexandre Keiji [UNIFESP]
dc.contributor.authorGrego, Kathleen Fernandes
dc.contributor.authorTanaka-Azevedo, Anita Mitico
dc.coverageSan Francisco
dc.date.accessioned2020-07-20T16:31:16Z
dc.date.available2020-07-20T16:31:16Z
dc.date.issued2018
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.en
dc.description.affiliationUniv Sao Paulo, Inst Pesquisas Tecnol, Inst Butantan, Interunidades Biotecnol, Sao Paulo, SP, Brazil
dc.description.affiliationInst Butantan, Lab Herpetol, Sao Paulo, SP, Brazil
dc.description.affiliationUniv Estadual Paulista, Inst Biociencias Litoral Paulista, Sao Vicente, SP, Brazil
dc.description.affiliationUniv Fed Sao Paulo, Dept Bioquim, Sao Paulo, SP, Brazil
dc.description.affiliationUnifespUniv Fed Sao Paulo, Dept Bioquim, Sao Paulo, SP, Brazil
dc.description.sourceWeb of Science
dc.description.sponsorshipFinanciadora de Estudos e Projetos and Fundacao de Amparo a Pesquisa do Estado de Sao Paulo - FAPESP
dc.description.sponsorshipCoordenacao de Aperfeicoamento de Pessoal de Nivel Superior-CAPES
dc.description.sponsorshipIDFAPESP: 2012/19321-9
dc.description.sponsorshipIDFAPESP: 2016/03839-0
dc.description.sponsorshipIDFAPESP: 2013/05357-4
dc.description.sponsorshipIDFAPESP: 2014/11108-0
dc.description.sponsorshipIDFAPESP: 2017/01890-0
dc.description.sponsorshipIDFAPESP: 2017/16908-2
dc.description.sponsorshipIDFAPESP: 2013/12826-0
dc.format.extent-
dc.identifierhttp://dx.doi.org/10.1371/journal.pone.0193105
dc.identifier.citationPlos One. San Francisco, v. 13, n. 3, 2018.
dc.identifier.doi10.1371/journal.pone.0193105
dc.identifier.fileWOS000426548200004.pdf
dc.identifier.issn1932-6203
dc.identifier.urihttps://repositorio.unifesp.br/handle/11600/55838
dc.identifier.wosWOS:000426548200004
dc.language.isoeng
dc.publisherPublic Library Science
dc.relation.ispartofPlos One
dc.rightsinfo:eu-repo/semantics/openAccess
dc.titlePurification and characterization of the first gamma-phospholipase inhibitor (gamma PLI) from Bothrops jararaca snake serumen
dc.typeinfo:eu-repo/semantics/article
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