A simple strategy for the purification of native recombinant full-length human RPL10 protein from inclusion bodies

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dc.contributor.authorPEREIRA, LARISSA M.
dc.contributor.authorSILVA, LUANA R.
dc.contributor.authorALVES, JOSEANE F.
dc.contributor.authorMARIN, NELIDA
dc.contributor.authorSILVA, FLAVIO S.
dc.contributor.authorMORGANTI, LIGIA
dc.contributor.authorSILVA, ISMAEL D.C.G.
dc.contributor.authorAFFONSO, REGINA
dc.coverageInternacionalpt_BR
dc.date.accessioned2014-12-23T12:43:27Z
dc.date.available2014-12-23T12:43:27Z
dc.date.issued2014pt_BR
dc.description.abstractThe L10 ribosomal protein (RPL10) plays a role in the binding of the 60 S and 40 S ribosomal subunits and in mRNA translation. The evidence indicates that RPL10 also has multiple extra-ribosomal functions, including tumor suppression. Recently, the presence of RPL10 in prostate and ovarian cancers was evaluated, and it was demonstrated to be associated with autistic disorders and premature ovarian failure. In the present work, we successfully cloned and expressed full-length human RPL10 (hRPL10) protein and isolated inclusion bodies containing this protein that had formed under mild growth conditions. The culture produced 376 mg of hRPL10 protein per liter of induced bacterial culture, of which 102.4 mg was present in the soluble fraction, and 25.6 mg was recovered at approximately 94% purity. These results were obtained using a two-step process of non-denaturing protein extraction from pelleted inclusion bodies. We studied the characteristics of this protein using circular dichroism spectroscopy and by monitoring the changes induced by the presence or absence of zinc ions using fluorescence spectrometry. The results demonstrated that the protein obtained using these non-conventional methods retained its secondary and tertiary structure. The conformational changes induced by the incorporation of zinc suggested that this protein could interact with Jun or the SH3 domain of c-yes. The results suggested that the strategy used to obtain hRPL10 is simple and could be applied to obtaining other proteins that are susceptible to degradation.
dc.format.extent115-120pt_BR
dc.identifier.citationPEREIRA, LARISSA M.; SILVA, LUANA R.; ALVES, JOSEANE F.; MARIN, NELIDA; SILVA, FLAVIO S.; MORGANTI, LIGIA; SILVA, ISMAEL D.C.G.; AFFONSO, REGINA. A simple strategy for the purification of native recombinant full-length human RPL10 protein from inclusion bodies. <b>Protein Expression and Purification</b>, v. 101, p. 115-120, 2014. DOI: <a href="https://dx.doi.org/10.1016/j.pep.2014.06.003">10.1016/j.pep.2014.06.003</a>. Disponível em: http://repositorio.ipen.br/handle/123456789/23190.
dc.identifier.doi10.1016/j.pep.2014.06.003
dc.identifier.issn1046-5928pt_BR
dc.identifier.orcidhttps://orcid.org/0000-0002-9264-4262
dc.identifier.orcidhttps://orcid.org/0000-0002-7870-1793
dc.identifier.urihttp://repositorio.ipen.br/handle/123456789/23190
dc.identifier.vol101pt_BR
dc.relation.ispartofProtein Expression and Purificationpt_BR
dc.rightsopenAccesspt_BR
dc.subjectribosomes
dc.subjectmessenger-rna
dc.subjectcloning
dc.subjectcarcinomas
dc.subjectgene therapy
dc.subjectinclusions
dc.subjectfolding model
dc.subjecttumor cells
dc.subjectinhibition
dc.subjectfluorescence spectroscopy
dc.titleA simple strategy for the purification of native recombinant full-length human RPL10 protein from inclusion bodiespt_BR
dc.typeArtigo de periódicopt_BR
dspace.entity.typePublication
ipen.autorJOSEANE FRANCISCA ALVES
ipen.autorFLAVIO SOUSA SILVA
ipen.autorREGINA AFFONSO
ipen.autorISMAEL DALE COTRIN GUERREIRO DA SILVA
ipen.autorLIGIA ELY MORGANTI FERREIRA DIAS
ipen.autorLUANA RISSI SILVA
ipen.autorLARISSA MIRANDA PEREIRA
ipen.codigoautor7554
ipen.codigoautor10312
ipen.codigoautor1547
ipen.codigoautor6994
ipen.codigoautor296
ipen.codigoautor3051
ipen.codigoautor6597
ipen.contributor.ipenauthorJOSEANE FRANCISCA ALVES
ipen.contributor.ipenauthorFLAVIO SOUSA SILVA
ipen.contributor.ipenauthorREGINA AFFONSO
ipen.contributor.ipenauthorISMAEL DALE COTRIN GUERREIRO DA SILVA
ipen.contributor.ipenauthorLIGIA ELY MORGANTI FERREIRA DIAS
ipen.contributor.ipenauthorLUANA RISSI SILVA
ipen.contributor.ipenauthorLARISSA MIRANDA PEREIRA
ipen.date.recebimento14-12pt_BR
ipen.identifier.fi1.695pt_BR
ipen.identifier.ipendoc20242pt_BR
ipen.identifier.iwosWoSpt_BR
ipen.identifier.ods3
ipen.range.fi1.500 - 2.999
ipen.type.genreArtigo
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sigepi.autor.atividadePEREIRA, LARISSA M.:6597:-1:S
sigepi.autor.atividadeSILVA, LUANA R.:3051:-1:N
sigepi.autor.atividadeALVES, JOSEANE F.:7554:-1:N
sigepi.autor.atividadeSILVA, FLAVIO S.:10312:820:N
sigepi.autor.atividadeMORGANTI, LIGIA:296:820:N
sigepi.autor.atividadeSILVA, ISMAEL D.C.G.:6994:-1:N
sigepi.autor.atividadeAFFONSO, REGINA:1547:820:N
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