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GST-tagged mouse estrogen receptor α-transactivation domain fusion protein is specifically degraded during its over-expression in E. coli and purification

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Abstract

Escherichia coli BL21 (DE3) is commonly used for the overproduction of fusion proteins. Using this system, we recently reported the overproduction of histidine-tagged mouse estrogen receptor (ER) α-ligand binding domain as an intact 30 kD protein and its inhibitory effect on the growth of bacteria. However, when GST-tagged mouse ERα transactivation domain (TAD) was overproduced using this system, it showed no effect on the growth of bacteria but was specifically degraded during its expression and purification. Here we report the expression of 47 kD GST-tagged mouse ERα-TAD protein, which was degraded partially and specifically into 46 and 43 kD fragments. This fusion protein was further degraded into 37, 31, 29 and 26 kD fragments during its purification by affinity chromatography. Such specific degradation of GST-tagged mouse ERα-TAD during its overproduction in E. coli and purification indicates the induction of specific protease and suggests the modification of expression system.

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Acknowledgments

The authors would like to thank Dr M.G. Parker, Imperial College, London, for providing mERα-TAD cDNA. Swati Ghosh is a recipient of Senior Research Fellowship from the University Grants Commission, India. This work was supported by grants from the Department of Biotechnology, Govt. of India (BT/PR3593/Med/14/468/2003) to M.K.T.

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  1. Biochemistry and Molecular Biology Laboratory, Department of Zoology, Banaras Hindu University, Varanasi, 221005, India

    M. K. Thakur & Swati Ghosh

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  1. M. K. Thakur
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  2. Swati Ghosh
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Correspondence to M. K. Thakur.

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Thakur, M.K., Ghosh, S. GST-tagged mouse estrogen receptor α-transactivation domain fusion protein is specifically degraded during its over-expression in E. coli and purification. Mol Biol Rep 37, 1335–1340 (2010). https://doi.org/10.1007/s11033-009-9512-8

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  • DOI: https://doi.org/10.1007/s11033-009-9512-8

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