Expression of eukaryotic proteins in Escherichia coli is challenging, especially when they contain disulfide bonds. Since the discovery of the prion protein (PrP) and its role in transmissible spongiform encephalopathies, the need to obtain large quantities of the recombinant protein for research purposes has been essential. Currently, production of recombinant PrP is achieved by refolding protocols. Here, we show that the co-expression of two different PrP with the human Quiescin Sulfhydryl OXidase (QSOX), a human chaperone with thiol/disulfide oxidase activity, in the cytoplasm of E. coli produces soluble recombinant PrP. The structural integrity of the soluble PrP has been confirmed by nuclear magnetic resonance spectroscopy, demonstrating that properly folded PrP can be easily expressed in bacteria. Furthermore, the soluble recombinant PrP produced with this method can be used for functional and structural studies

A novel expression system for production of soluble prion proteins in E.coli / Abskharon, Rn; Ramboarina, S; EL HASSAN, H; Gad, W; Apostol, Mi; Giachin, G; Legname, Giuseppe; Steyaert, J; Messens, J; Soror, Sh; Wohlkonig, A.. - In: MICROBIAL CELL FACTORIES. - ISSN 1475-2859. - 11:10 Juanuary(2012), pp. 1-11. [10.1186/1475-2859-11-6]

A novel expression system for production of soluble prion proteins in E.coli

GIACHIN G;Legname, Giuseppe;
2012-01-01

Abstract

Expression of eukaryotic proteins in Escherichia coli is challenging, especially when they contain disulfide bonds. Since the discovery of the prion protein (PrP) and its role in transmissible spongiform encephalopathies, the need to obtain large quantities of the recombinant protein for research purposes has been essential. Currently, production of recombinant PrP is achieved by refolding protocols. Here, we show that the co-expression of two different PrP with the human Quiescin Sulfhydryl OXidase (QSOX), a human chaperone with thiol/disulfide oxidase activity, in the cytoplasm of E. coli produces soluble recombinant PrP. The structural integrity of the soluble PrP has been confirmed by nuclear magnetic resonance spectroscopy, demonstrating that properly folded PrP can be easily expressed in bacteria. Furthermore, the soluble recombinant PrP produced with this method can be used for functional and structural studies
2012
11
10 Juanuary
1
11
6
Abskharon, Rn; Ramboarina, S; EL HASSAN, H; Gad, W; Apostol, Mi; Giachin, G; Legname, Giuseppe; Steyaert, J; Messens, J; Soror, Sh; Wohlkonig, A.
File in questo prodotto:
File Dimensione Formato  
Abskharon_MicrobCellFact_2012.pdf

accesso aperto

Descrizione: Open Access article
Tipologia: Versione Editoriale (PDF)
Licenza: Creative commons
Dimensione 2.86 MB
Formato Adobe PDF
2.86 MB Adobe PDF Visualizza/Apri

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/20.500.11767/14662
Citazioni
  • ???jsp.display-item.citation.pmc??? 11
  • Scopus 24
  • ???jsp.display-item.citation.isi??? 22
social impact