A simple strategy for the purification of native recombinant full-length human RPL10 protein from inclusion bodies
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Protein Expression and Purification
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The 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.
Como referenciar
PEREIRA, 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. Protein Expression and Purification, v. 101, p. 115-120, 2014. DOI: 10.1016/j.pep.2014.06.003. Disponível em: http://repositorio.ipen.br/handle/123456789/23190. Acesso em: 30 Dec 2025.
Esta referência é gerada automaticamente de acordo com as normas do estilo IPEN/SP (ABNT NBR 6023) e recomenda-se uma verificação final e ajustes caso necessário.