DogCatcher allows loop-friendly protein-protein ligation
Journal article
Authors | Heeble, A. H., Yadav, VK, Ferla, M. P., Bauer, C. C., Chuntharpursat-Bon, C., Huang, J., Bon, R. S. and Howarth, M. |
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Abstract | There are many efficient ways to connect proteins at termini. However, connecting at a loop is difficult because of lower flexibility and variable environment. Here, we have developed DogCatcher, a protein that forms a spontaneous isopeptide bond with DogTag peptide. DogTag/DogCatcher was generated initially by splitting a Streptococcus pneumoniae adhesin. We optimized DogTag/DogCatcher through rational design and evolution, increasing reaction rate by 250-fold and establishing millimolar solubility of DogCatcher. When fused to a protein terminus, DogTag/DogCatcher reacts slower than SpyTag003/SpyCatcher003. However, inserted in loops of a fluorescent protein or enzyme, DogTag reacts much faster |
Keywords | Protein engineering; isopeptide bond; covalent labeling; Ligation |
Year | 2022 |
Journal | Cell Chemical Biology |
Journal citation | 29 (2), p. 339–350 |
Publisher | Elsevier |
ISSN | 2451-9448 |
Digital Object Identifier (DOI) | https://doi.org/10.1016/j.chembiol.2021.07.005 |
Web address (URL) | https://www.cell.com/cell-chemical-biology/fulltext/S2451-9456(21)00315-9?_returnURL=https%3A%2F%2Flinkinghub.elsevier.com%2Fretrieve%2Fpii%2FS2451945621003159%3Fshowall%3Dtrue |
https://pubmed.ncbi.nlm.nih.gov/34324879/ | |
Accepted author manuscript | File Access Level Open |
Publisher's version | License File Access Level Open |
Output status | Published |
Publication dates | |
Online | 28 Jul 2021 |
Publication process dates | |
Deposited | 16 Dec 2022 |
https://repository.derby.ac.uk/item/9vqy3/dogcatcher-allows-loop-friendly-protein-protein-ligation
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