A novel expression system for production of soluble prion proteins in E. coli
<p>Abstract</p> <p>Expression of eukaryotic proteins in <it>Escherichia coli </it>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 l...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2012-01-01
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| Series: | Microbial Cell Factories |
| Online Access: | http://www.microbialcellfactories.com/content/11/1/6 |
| _version_ | 1811249634009415680 |
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| author | Abskharon Romany NN Ramboarina Stephanie El Hassan Hassan Gad Wael Apostol Marcin I Giachin Gabriele Legname Giuseppe Steyaert Jan Messens Joris Soror Sameh H Wohlkonig Alexandre |
| author_facet | Abskharon Romany NN Ramboarina Stephanie El Hassan Hassan Gad Wael Apostol Marcin I Giachin Gabriele Legname Giuseppe Steyaert Jan Messens Joris Soror Sameh H Wohlkonig Alexandre |
| author_sort | Abskharon Romany NN |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Expression of eukaryotic proteins in <it>Escherichia coli </it>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 <it>E. coli </it>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.</p> |
| first_indexed | 2024-04-12T15:51:05Z |
| format | Article |
| id | doaj.art-c6963cfd365f4ef090b25ac4c0153070 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-04-12T15:51:05Z |
| publishDate | 2012-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-c6963cfd365f4ef090b25ac4c01530702022-12-22T03:26:31ZengBMCMicrobial Cell Factories1475-28592012-01-01111610.1186/1475-2859-11-6A novel expression system for production of soluble prion proteins in E. coliAbskharon Romany NNRamboarina StephanieEl Hassan HassanGad WaelApostol Marcin IGiachin GabrieleLegname GiuseppeSteyaert JanMessens JorisSoror Sameh HWohlkonig Alexandre<p>Abstract</p> <p>Expression of eukaryotic proteins in <it>Escherichia coli </it>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 <it>E. coli </it>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.</p>http://www.microbialcellfactories.com/content/11/1/6 |
| spellingShingle | Abskharon Romany NN Ramboarina Stephanie El Hassan Hassan Gad Wael Apostol Marcin I Giachin Gabriele Legname Giuseppe Steyaert Jan Messens Joris Soror Sameh H Wohlkonig Alexandre A novel expression system for production of soluble prion proteins in E. coli Microbial Cell Factories |
| title | A novel expression system for production of soluble prion proteins in E. coli |
| title_full | A novel expression system for production of soluble prion proteins in E. coli |
| title_fullStr | A novel expression system for production of soluble prion proteins in E. coli |
| title_full_unstemmed | A novel expression system for production of soluble prion proteins in E. coli |
| title_short | A novel expression system for production of soluble prion proteins in E. coli |
| title_sort | novel expression system for production of soluble prion proteins in e coli |
| url | http://www.microbialcellfactories.com/content/11/1/6 |
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