Solubility of recombinant Src homology 2 domains expressed in E. colican be predicted by TANGO

BMC Biotechnology

Research Article

Thorny Cecilie Bie Andersen¹  Cecilie Dahl Hem¹  Anne Spurkland¹  Kjersti Lindsjø¹  Lise Koll¹  Amy H Andreotti²  Per Eugen Kristiansen³  Lars Skjeldal⁴ 

[1] Department of Anatomy, Institute of Basal Medical Sciences, University of Oslo, Oslo, Norway;Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, Iowa, USA;Department of Biosciences, University of Oslo, Oslo, Norway;Department of Chemistry, Biochemistry and Food science, Norwegian University of Life Sciences, Ås, Norway;

关键词: Bacterial inclusion bodies;    Protein aggregation;    Recombinant protein expression;    SH2 domain;    SH2D2A;    Protein solubility;   

DOI  :  10.1186/1472-6750-14-3

 received in 2013-03-26, accepted in 2014-01-07,  发布年份 2014

来源: Springer

【 摘 要 】

BackgroundSignalling proteins often contain several well defined and conserved protein domains. Structural analyses of such domains by nuclear magnetic spectroscopy or X-ray crystallography may greatly inform the function of proteins. A limiting step is often the production of sufficient amounts of the recombinant protein. However, there is no particular way to predict whether a protein will be soluble when expressed in E.coli. Here we report our experience with expression of a Src homology 2 (SH2) domain.ResultsThe SH2 domain of the SH2D2A protein (or T cell specific adapter protein, TSAd) forms insoluble aggregates when expressed as various GST-fusion proteins in Escherichia coli (E. coli). Alteration of the flanking sequences, or growth temperature influenced expression and solubility of TSAd-SH2, however overall yield of soluble protein remained low. The algorithm TANGO, which predicts amyloid fibril formation in eukaryotic cells, identified a hydrophobic sequence within the TSAd-SH2 domain with high propensity for beta-aggregation. Mutation to the corresponding amino acids of the related HSH2- (or ALX) SH2 domain increased the yield of soluble TSAd-SH2 domains. High beta-aggregation values predicted by TANGO correlated with low solubility of recombinant SH2 domains as reported in the literature.ConclusionsSolubility of recombinant proteins expressed in E.coli can be predicted by TANGO, an algorithm developed to determine the aggregation propensity of peptides. Targeted mutations representing corresponding amino acids in similar protein domains may increase solubility of recombinant proteins.

【 授权许可】

Unknown   
© Andersen et al.; licensee BioMed Central Ltd. 2014. This article is published under license to BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.