Biochemical analyses of proteins from Duttaphrynus melanostictus (Bufo melanostictus) skin secretion: soluble protein retrieval from a viscous matrix by ion-exchange batch sample preparation

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dc.contributorLaboratório de Bioquímica e Biofísicapt_BR
dc.contributor.authorMariano, Douglas Oscar Ceolinpt_BR
dc.contributor.authorMessias, Marcela Di Giacomopt_BR
dc.contributor.authorPrezotto-Neto, José Pedropt_BR
dc.contributor.authorPimenta, Daniel Carvalhopt_BR
dc.contributor.authorSpencer, Patrick J.pt_BR
dc.date.accessioned2020-07-09T21:21:02Z-
dc.date.available2020-07-09T21:21:02Z-
dc.date.issued2018pt_BR
dc.identifier.citationMariano DOC, Messias MDG, Prezotto-Neto JP, Pimenta DC, Spencer PJ.. Biochemical analyses of proteins from Duttaphrynus melanostictus (Bufo melanostictus) skin secretion: soluble protein retrieval from a viscous matrix by ion-exchange batch sample preparation. Protein J. 2018 Aug;37(4):380–89. doi:10.1007/s10930-018-9780-z.pt_BR
dc.identifier.urihttps://repositorio.butantan.gov.br/handle/butantan/2539-
dc.description.abstractA crucial step in scientific analysis can be sample preparation, and its importance increases in the same rate as the sensitivity of the following employed/desired analytical technique does. The need to analyze complex, viscous matrices is not new, and diverse approaches have been employed, with different success rates depending on the intended molecules. Solid-phase extraction, for example, has been successfully used in sample preparation for organic molecules and peptides. However, due to the usual methodological conditions, biologically active proteins are not successfully retrieved by this technique, resulting in a low rate of protein identification reported for the viscous amphibian skin secretion. Here we describe an ion-exchange batch processing sample preparation technique that allows viscous or adhesive materials (as some amphibian skin secretions) to be further processed by classical liquid chromatography approaches. According to our protocol, samples were allowed to equilibrate with a specific resin that was washed with appropriated buffers in order to yield the soluble protein fraction. In order to show the efficiency of our methodology, we have compared our results to classically prepared skin secretion, i.e., by means of filtration and centrifugation. After batch sample preparation, we were able to obtain reproductive resolved protein chromatographic profiles, as revealed by SDS-PAGE, and retrieve some biological activities, namely, hydrolases belonging to serine peptidase family. Not only that, but also the unbound fraction was rich in low molecular mass molecules, such as alkaloids and steroids, making this sample preparation technique also suitable for the enrichment of such molecules.pt_BR
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)pt_BR
dc.description.sponsorshipFinanciadora de Estudos e Projetos (FINEP)pt_BR
dc.description.sponsorshipCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)pt_BR
dc.format.extentp. 380-389pt_BR
dc.language.isoEnglishpt_BR
dc.relation.ispartofThe Protein Journalpt_BR
dc.titleBiochemical analyses of proteins from Duttaphrynus melanostictus (Bufo melanostictus) skin secretion: soluble protein retrieval from a viscous matrix by ion-exchange batch sample preparationpt_BR
dc.typeArticlept_BR
dc.identifier.doi10.1007/s10930-018-9780-zpt_BR
dc.identifier.urlhttps://doi.org/10.1007/s10930-018-9780-zpt_BR
dc.contributor.externalInstituto de Pesquisas Energéticas e Nucleares (IPEN)¦¦Brasilpt_BR
dc.identifier.citationvolume37pt_BR
dc.identifier.citationissue4pt_BR
dc.subject.keywordBatch chromatographypt_BR
dc.subject.keywordIon-exchange chromatographypt_BR
dc.subject.keywordViscous samplept_BR
dc.subject.keywordAmphibian skin secretionpt_BR
dc.subject.keywordHydrolasept_BR
dc.relation.ispartofabbreviatedProtein Jpt_BR
dc.identifier.citationabntv. 37, n. 4, p. 380–89, ago. 2018pt_BR
dc.identifier.citationvancouver2018 Aug;37(4):380–89pt_BR
dc.contributor.butantanPimenta, Daniel Carvalho|:Pesquisador:Docente Permanente PPGTOX|:Laboratório de Bioquímica e Biofísica|:Autor de correspondênciapt_BR
dc.contributor.butantanMariano, Douglas Oscar Ceolin|:Aluno|:Laboratório de Bioquímica e Biofísica|:PrimeiroAutorpt_BR
dc.sponsorship.butantanConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)¦¦303792/2016-7pt_BR
dc.sponsorship.butantanCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)¦¦969130pt_BR
dc.sponsorship.butantanFinanciadora de Estudos e Projetos (FINEP)¦¦01.09.0278.04pt_BR
dc.identifier.bvsccBR78.1pt_BR
dc.identifier.bvsdbIBProdpt_BR
dc.description.dbindexedYespt_BR
item.fulltextSem Texto completo-
item.grantfulltextnone-
item.languageiso639-1English-
item.openairetypeArticle-
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