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一种在大肠杆菌中生产可溶性朊病毒蛋白的新型表达系统。

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

机构信息

VIB, Department of Structural Biology, Brussels, Belgium.

出版信息

Microb Cell Fact. 2012 Jan 10;11:6. doi: 10.1186/1475-2859-11-6.

DOI:10.1186/1475-2859-11-6
PMID:22233534
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3283519/
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.

摘要

真核蛋白在大肠杆菌中的表达具有挑战性，尤其是当它们含有二硫键时。自从发现朊病毒蛋白（PrP）及其在传染性海绵状脑病中的作用以来，为了研究目的获得大量重组蛋白已变得至关重要。目前，通过重折叠方案来生产重组 PrP。在这里，我们表明，在大肠杆菌细胞质中共表达两种不同的 PrP 和人类 Q 硫氧还蛋白（QSOX），一种具有巯基/二硫键氧化酶活性的人类伴侣蛋白，可以产生可溶性重组 PrP。通过核磁共振波谱法证实了可溶性 PrP 的结构完整性，表明正确折叠的 PrP 可以在细菌中轻松表达。此外，用这种方法生产的可溶性重组 PrP 可用于功能和结构研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7ec/3283519/b2ac0a70dcfa/1475-2859-11-6-10.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7ec/3283519/3780fb7a6544/1475-2859-11-6-8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7ec/3283519/95cc355c3b74/1475-2859-11-6-9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7ec/3283519/b2ac0a70dcfa/1475-2859-11-6-10.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7ec/3283519/c79cd7df0e7e/1475-2859-11-6-1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7ec/3283519/34de8ec90f7b/1475-2859-11-6-2.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7ec/3283519/9eac3aba5168/1475-2859-11-6-4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7ec/3283519/226270f1569e/1475-2859-11-6-5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7ec/3283519/e09d176e37cd/1475-2859-11-6-6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7ec/3283519/4bc0c94bfa73/1475-2859-11-6-7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7ec/3283519/3780fb7a6544/1475-2859-11-6-8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7ec/3283519/95cc355c3b74/1475-2859-11-6-9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7ec/3283519/b2ac0a70dcfa/1475-2859-11-6-10.jpg

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