Hatlem, D., Trunk, T., Linke, D. and Leo, J.C. ORCID: 0000-0002-7066-7527, 2019. Catching a SPY: using the SpyCatcher-SpyTag and related systems for labeling and localizing bacterial proteins. International Journal of Molecular Sciences, 20 (9): 2129. ISSN 1661-6596
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Abstract
The SpyCatcher-SpyTag system was developed seven years ago as a method for protein ligation. It is based on a modified domain from a Streptococcus pyogenes surface protein (SpyCatcher), which recognizes a cognate 13-amino-acid peptide (SpyTag). Upon recognition, the two form a covalent isopeptide bond between the side chains of a lysine in SpyCatcher and an aspartate in SpyTag. This technology has been used, among other applications, to create covalently stabilized multi-protein complexes, for modular vaccine production, and to label proteins (e.g., for microscopy). The SpyTag system is versatile as the tag is a short, unfolded peptide that can be genetically fused to exposed positions in target proteins; similarly, SpyCatcher can be fused to reporter proteins such as GFP, and to epitope or purification tags. Additionally, an orthogonal system called SnoopTag-SnoopCatcher has been developed from an S. pneumoniae pilin that can be combined with SpyCatcher-SpyTag to produce protein fusions with multiple components. Furthermore, tripartite applications have been produced from both systems allowing the fusion of two peptides by a separate, catalytically active protein unit, SpyLigase or SnoopLigase. Here, we review the current state of the SpyCatcher-SpyTag and related technologies, with a particular emphasis on their use in vaccine development and in determining outer membrane protein localization and topology of surface proteins in bacteria.
Item Type: | Journal article | ||||
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Description: | In special issue: Microbial Virulence Factors. | ||||
Publication Title: | International Journal of Molecular Sciences | ||||
Creators: | Hatlem, D., Trunk, T., Linke, D. and Leo, J.C. | ||||
Publisher: | MDPI | ||||
Date: | 1 May 2019 | ||||
Volume: | 20 | ||||
Number: | 9 | ||||
ISSN: | 1661-6596 | ||||
Identifiers: |
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Rights: | © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). | ||||
Divisions: | Schools > School of Science and Technology | ||||
Record created by: | Jill Tomkinson | ||||
Date Added: | 18 Jun 2019 13:12 | ||||
Last Modified: | 25 Jun 2019 12:54 | ||||
URI: | https://irep.ntu.ac.uk/id/eprint/36841 |
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