 visitante ::
identificación
|
|||||||||||||||
|
Buscar | Enviar | Ayuda | Servicio de Bibliotecas | Sobre el DDD | Català English Español | |||||||||
| Página principal > Artículos > Artículos publicados > Co-production of GroELS discriminates between intrinsic and thermally-induced recombinant protein aggregation during substrate quality control |
(Centro de Investigación Biomédica en Red de Bioingeniería, Biomateriales y Nanomedicina) (Universitat Autònoma de Barcelona. Institut de Biotecnologia i de Biomedicina "Vicent Villar Palasí") (Universitat Autònoma de Barcelona. Institut de Biotecnologia i de Biomedicina "Vicent Villar Palasí")| Fecha: | 2011 |
| Resumen: | The effects and effectiveness of the chaperone pair GroELS on the yield and quality of recombinant polypeptides produced in Escherichia coli are matter of controversy, as the reported activities of this complex are not always consistent and eventually indicate undesired side effects. The divergence in the reported data could be due, at least partially, to different experimental conditions in independent research approaches. We have then selected two structurally different model proteins (namely GFP and E. coli β-galactosidase) and two derived aggregation-prone fusions to explore, in a systematic way, the eventual effects of GroELS co-production on yield, solubility and conformational quality. Host cells were cultured at two alternative temperatures below the threshold at which thermal stress is expected to be triggered, to minimize the involvement of independent stress factors. From the analysis of protein yield, solubility and biological activity of the four model proteins produced alone or along the chaperones, we conclude that GroELS impacts on yield and quality of aggregation-prone proteins with intrinsic determinants but not on thermally induced protein aggregation. No effective modifications of protein solubility have been observed, but significant stabilization of small (encapsulable) substrates and moderate chaperone-induced degradation of larger (excluded) polypeptides. These findings indicate that the activities of this chaperone pair in the context of actively producing recombinant bacteria discriminate between intrinsic and thermally-induced protein aggregation, and that the side effects of GroELS overproduction might be determined by substrate size. |
| Ayudas: | Ministerio de Ciencia e Innovación BFU2010/17450 Ministerio de Ciencia e Innovación ACI2009/0919 Ministerio de Ciencia e Innovación EUI2008/03610 Agència de Gestió d'Ajuts Universitaris i de Recerca 2009/SGR-108 |
| Nota: | Altres ajuts: ERANET-UB98-007 |
| Derechos: | Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.  |
| Lengua: | Anglès |
| Documento: | Article ; recerca ; Versió publicada |
| Materia: | Protein Yield ; Recombinant Protein Production ; Recombinant Polypeptide ; Conformational Quality ; Recombinant Protein Yield |
| Publicado en: | Microbial cell factories, Vol. 10 (October 2011) , art. 79, ISSN 1475-2859 |
