The DOI matches its Crossref record; 3 of the 3 references it cites were found in a registry or an index.
Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin
Open accessAbstract
Protein interactions with peptides generally have low thermodynamic and mechanical stability. Streptococcus pyogenes fibronectin-binding protein FbaB contains a domain with a spontaneous isopeptide bond between Lys and Asp. By splitting this domain and rational engineering of the fragments, we obtained a peptide (SpyTag) which formed an amide bond to its protein partner (SpyCatcher) in minutes. Reaction occurred in high yield simply upon mixing and amidst diverse conditions of pH, temperature, and buffer. SpyTag could be fused at either terminus or internally and reacted specifically at the mammalian cell surface. Peptide binding was not reversed by boiling or competing peptide. Single-molecule dynamic force spectroscopy showed that SpyTag did not separate from SpyCatcher until the force exceeded 1 nN, where covalent bonds snap. The robust reaction conditions and irreversible linkage of SpyTag shed light on spontaneous isopeptide bond formation and should provide a targetable lock in cells and a stable module for new protein architectures.
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Cites
Global analysis of protein localization in budding yeast
Huh, W.-K., Falvo, J. V., Gerke, L. C., Carroll, A. S., Howson, R. W., Weissman, J. S., & O'Shea, E. K. (2003) · Nature · doi:10.1038/nature02026 · Cited by 4,248 · cited 1×
“Peptides are very useful minimally disruptive probes (5) but they are also “slippery”- antibodies or other proteins typically bind peptides with low affinity and poor mechanical strength (6–9).”
BackgroundThe Design of Coiled-Coil Structures and Assemblies
Woolfson, D. N. (2005) · Advances in Protein Chemistry · doi:10.1016/s0065-3233(05)70004-8 · Cited by 585
Protein sorting to the cell wall envelope of Gram-positive bacteria
Ton-That, H., Marraffini, L. A., & Schneewind, O. (2004) · Biochimica et Biophysica Acta (BBA) - Molecular Cell Research · doi:10.1016/j.bbamcr.2004.04.014 · Cited by 266 · cited 1×
“With this split system we provide the clearest evidence that reaction can occur in minutes, consistent with the biological time scale for export of these proteins (13, 26).”
Background
Cited by
Plasticity in binding confers selectivity in ligand-induced protein degradation
Nowak, R. P., DeAngelo, S. L., Buckley, D., He, Z., Donovan, K. A., An, J., Safaee, N., Jedrychowski, M. P., Ponthier, C. M., Ishoey, M., et al. (2018) · Nature Chemical Biology · doi:10.1038/s41589-018-0055-y · Cited by 615
Major findings and recent advances in virus–like particle (VLP)-based vaccines
Mohsen, M. O., Zha, L., Cabral-Miranda, G., & Bachmann, M. F. (2017) · Seminars in Immunology · doi:10.1016/j.smim.2017.08.014 · Cited by 555
Donovan, K. A., An, J., Nowak, R. P., Yuan, J. C., Fink, E. C., Berry, B. C., Ebert, B. L., & Fischer, E. S. (2018) · eLife · doi:10.7554/elife.38430 · Cited by 509
“Spycatcher (Zakeri et al., 2012) containing a Ser50Cys mutation was obtained as a synthetic dsDNA fragment from IDT (Integrated DNA technologies) and subcloned as a GST-TEV fusion protein in a pET-Duet-derived vector.”
MethodCheng, K., Zhao, R., Li, Y., Qi, Y., Wang, Y., Zhang, Y., Qin, H., Qin, Y., Chen, L., Li, C., et al. (2021) · Nature Communications · doi:10.1038/s41467-021-22308-8 · Cited by 480
Switchable Fluorophores for Single-Molecule Localization Microscopy
Li, H., & Vaughan, J. C. (2018) · Chemical Reviews · doi:10.1021/acs.chemrev.7b00767 · Cited by 319
Metadata
- Authors
- Zakeri, B., Fierer, J. O., Celik, E., Chittock, E. C., Schwarz-Linek, U., Moy, V. T., & Howarth, M.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 2012-02-24
- Volume
- 109
- Issue
- 12
- Pages
- E690-7
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W1976672416
- Cited by
- 1,662
- References
- 3
- FWCI
- 6.04
- Type
- journal article
- Language
- en
- Open access
- green
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- Zakeri, B., Fierer, J. O., Celik, E., Chittock, E. C., Schwarz-Linek, U., Moy, V. T., & Howarth, M. (2012). Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.1115485109
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.
