dc.contributor.authorGoh, Hui Chin
dc.contributor.authorSobota, Radoslaw M.
dc.contributor.authorGhadessy, Farid J.
dc.contributor.authorNirantar, Saurabh
dc.identifier.citationGoh, H. C., Sobota, R. M., Ghadessy, F. J.,& Nirantar, S. (2017). Going native: Complete removal of protein purification affinity tags by simple modification of existing tags and proteases. Protein Expression and Purification, 12918-24.en_US
dc.description.abstractProtein purification typically involves expressing a recombinant gene comprising a target protein fused to a suitable affinity tag. After purification, it is often desirable to remove the affinity tag to prevent interference with downstream functions of the target protein. This is mainly accomplished by placing a protease site between the tag and the target protein. Typically, a small oligopeptide ‘stub’ C-terminal to the cleavage site remains attached to the target protein due to the requirements of sequence-specific proteases. Furthermore, steric hindrance can also limit protease efficiency. Here, we show that respectively fusing the interacting ePDZ-b/ARVCF protein-peptide pair to the target protein and a protease enables efficient processing of a minimised sequence comprising only residues N-terminal to the cleavage site. Interaction of the protein-peptide pair enforces proximity of the protease and its minimised cleavage sequence, enhancing both catalysis of a sub-optimal site and overcoming steric hindrance. This facilitates the high yield purification of fully native target proteins without recourse to specialised purification columns.en_US
dc.description.sponsorshipNMRC (Natl Medical Research Council, S’pore)en_US
dc.format.extent24 p.en_US
dc.relation.ispartofseriesProtein Expression and Purificationen_US
dc.rights© 2016 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).en_US
dc.subjectProtein purificationen_US
dc.subjectAffinity tagsen_US
dc.titleGoing native: Complete removal of protein purification affinity tags by simple modification of existing tags and proteasesen_US
dc.typeJournal Article
dc.contributor.schoolSchool of Biological Sciencesen_US
dc.description.versionPublished versionen_US

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