An efficient system for small protein expression and refolding Journal Article
| Authors: | Cheng, Y.; Patel, D. J. |
| Article Title: | An efficient system for small protein expression and refolding |
| Abstract: | The low expression yield and poor refolding efficiency of small recombinant proteins expressed in Escherichia coli have continued to hinder the large-scale purification of such proteins for structural and biological investigations. A system based on a small fusion partner, the B1 domain of Streptococcal protein G (GB1), was utilized to overcome this problem. We have tested this system on a small cysteine-rich toxin, mutant myotoxin α (MyoP20G). The highly expressed fusion protein was refolded using an unfolding/refolding protocol. Due to the small size of GB1, we were able to monitor the unfolding/refolding status by heteronuclear single quantum coherence (HSQC) NMR spectroscopy. The final product yielded well-resolved NMR spectra, with a topology corresponding to the natural product. We conclude that GB1 not only increases the expression level but also enhances the refolding of small proteins. © 2004 Elsevier Inc. All rights reserved. |
| Keywords: | protein expression; nonhuman; bacterial proteins; recombinant fusion proteins; feasibility studies; protein purification; escherichia coli; recombinant protein; magnetic resonance spectroscopy; nuclear magnetic resonance spectroscopy; protein folding; protein structure; natural product; molecular weight; cysteine; productivity; mutant; gene expression regulation, bacterial; gene transfer techniques; purification; streptococcus; toxin; protein engineering; genetic enhancement; nmr spectroscopy; protein g; priority journal; article; protein refolding; crotalid venoms |
| Journal Title: | Biochemical and Biophysical Research Communications |
| Volume: | 317 |
| Issue: | 2 |
| ISSN: | 0006-291X |
| Publisher: | Elsevier Science, Inc. |
| Date Published: | 2004-04-30 |
| Start Page: | 401 |
| End Page: | 405 |
| Language: | English |
| DOI: | 10.1016/j.bbrc.2004.03.068 |
| PROVIDER: | scopus |
| PUBMED: | 15063772 |
| PMCID: | PMC4693640 |
| DOI/URL: | |
| Notes: | Erratum/Corrigendum issued, see DOI: 10.1016/j.bbrc.2005.06.157 -- Biochem. Biophys. Res. Commun. -- Cited By (since 1996):21 -- Export Date: 16 June 2014 -- CODEN: BBRCA -- Source: Scopus |
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