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

Full metadata record
DC FieldValueLanguage
dc.contributorLab. Bioquímicapt_BR
dc.contributorPrograma de Pós-Graduação em Ciências – Toxinologia (PPGTox)pt_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.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.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.sponsorship(CNPq) Conselho Nacional de Desenvolvimento Científico e Tecnológicopt_BR
dc.description.sponsorship(FINEP) Financiadora de Estudos e Projetospt_BR
dc.description.sponsorship(CAPES) Coordenação de Aperfeiçoamento de Pessoal de Nível Superiorpt_BR
dc.format.extentp. 380-389pt_BR
dc.relation.ispartofThe Protein journalpt_BR
dc.rightsRestricted accesspt_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.contributor.external(IPEN) Instituto de Pesquisas Energéticas e Nuclearespt_BR
dc.subject.keywordbatch chromatographypt_BR
dc.subject.keywordion-exchange chromatographypt_BR
dc.subject.keywordviscous samplept_BR
dc.subject.keywordamphibian skin secretionpt_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|:Lab. Bioquímica|:Autor de correspondênciapt_BR
dc.contributor.butantanMariano, Douglas Oscar Ceolin|:Aluno|:Lab. Bioquímica|:PrimeiroAutorpt_BR
dc.sponsorship.butantan(CNPq) Conselho Nacional de Desenvolvimento Científico e Tecnológico¦¦303792/2016-7pt_BR
dc.sponsorship.butantan(CAPES) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior¦¦969130pt_BR
dc.sponsorship.butantanFinanciadora de Estudos e Projetos (FINEP)¦¦01.09.0278.04pt_BR
item.fulltextCom Texto completo-
Appears in Collections:Artigos

Show simple item record

The access to the publications deposited in this repository respects the licenses from journals and publishers.