Enhancing recombinant protein solubility with ubiquitin-like small archeal modifying protein fusion partners

Journal of Microbiological Methods
Sándor VargaIstván Nagy

Abstract

A variety of protein expression tags with different biochemical properties has been used to enhance the yield and solubility of recombinant proteins. Ubiquitin, SUMO (small ubiquitin-like modifier) and prokaryotic ubiquitin like MoaD (molybdopterin synthase, small subunit) fusion tags are getting more popular because of their small size. In this paper we report on the use of ubiquitin-like small archaeal modifier proteins (SAMPs) as fusion tags since they proved to increase expression yield, stability and solubility in our experiments. Equally important, they did not co-purify with proteins of the expression host and there was information that their specific JAB1/MPN/Mov34 metalloenzyme (JAMM) protease can recognize the C-terminal VSGG sequence when SAMPs fused, either branched or linearly to target proteins, and cleave it specifically. SAMPs and JAMM proteases from Haloferax volcanii, Thermoplasma acidophilum, Methanococcoides burtonii and Nitrosopumilus maritimus were selected, cloned, expressed heterologously in Escherichia coli and tested as fusion tags and cleaving proteases, respectively. Investigated SAMPs enhanced protein expression and solubility on a wide scale. T. acidophilum SAMPs Ta0895 and Ta01019 were the best pe...Continue Reading

References

Dec 21, 2004·Journal of Biotechnology·Hans Peter Sørensen, Kim Kusk Mortensen
Jan 24, 2006·Protein Expression and Purification·José ArnauJohn Pedersen
Jun 20, 2006·Current Opinion in Biotechnology·Dominic Esposito, Deb K Chatterjee
Aug 30, 2011·Protein Expression and Purification·David S Waugh
Nov 13, 2012·Trends in Microbiology·Julie A Maupin-Furlow
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Feb 12, 2014·Proceedings of the National Academy of Sciences of the United States of America·Ganesh Ramnath PathareWolfgang Baumeister
May 27, 2014·Frontiers in Microbiology·Germán L Rosano, Eduardo A Ceccarelli

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