Journal
PROTEIN EXPRESSION AND PURIFICATION
Volume 116, Issue -, Pages 42-49Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.pep.2015.08.019
Keywords
Fusion proteins; Solubility enhancement tag; SUMO system; Ulp1 protease; TonB; Scytovirin; Protein ligation
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Funding
- Academy of Finland [137995]
- Sigrid Juselius Foundation
- National Doctoral Programme in Informational and Structural Biology (ISB)
- Biocenter Finland
- Academy of Finland (AKA) [137995, 137995] Funding Source: Academy of Finland (AKA)
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Availability of highly purified proteins in quantity is crucial for detailed biochemical and structural investigations. Fusion tags are versatile tools to facilitate efficient protein purification and to improve soluble overexpression of proteins. Various purification and fusion tags have been widely used for overexpression in Escherichia coli. However, these tags might interfere with biological functions and/or structural investigations of the protein of interest. Therefore, an additional purification step to remove fusion tags by proteolytic digestion might be required. Here, we describe a set of new vectors in which yeast SUMO (SMT3) was used as the highly specific recognition sequence of ubiquitin-like protease 1, together with other commonly used solubility enhancing proteins, such as glutathione S-transferase, maltose binding protein, thioredoxin and trigger factor for optimizing soluble expression of protein of interest. This tandem SUMO (T-SUMO) fusion system was tested for soluble expression of the C-terminal domain of TonB from different organisms and for the antiviral protein scytovirin. (C) 2015 Elsevier Inc. All rights reserved.
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