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Fluorescent-conjugated polymer superquenching facilitates highly sensitive detection of proteases
Open accessAbstract
Sensor formats have been developed for detecting the activity of proteolytic enzymes based on fluorescent conjugated polymer superquenching. These sensors employ a reactive peptide sequence within a tether linking a quencher to a biotin. The peptide binds to sensors containing colocated biotin-binding protein and fluorescent polymer by means of biotin–biotin binding protein interactions, resulting in a strong quenching of polymer fluorescence. Enzyme-mediated cleavage of the peptide results in a reversal of the fluorescence quenching. These assays for protease activity are simple, sensitive, fast, and have the specificity required for screening chemical libraries for novel protease inhibitors in a high-throughput screening assay environment. These assays have been demonstrated for enterokinase, caspase-3/7, and β-secretase.
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Kang, J., Lemaire, H.-G., Unterbeck, A., Salbaum, J. M., Masters, C. L., Grzeschik, K.-H., Multhaup, G., Beyreuther, K., & Müller-Hill, B. (1987) · Nature · doi:10.1038/325733a0 · Cited by 4,619
Vassar, R., Bennett, B. D., Babu-Khan, S., Kahn, S., Mendiaz, E. A., Denis, P., Teplow, D. B., Ross, S., Amarante, P., Loeloff, R., et al. (1999) · Science · doi:10.1126/science.286.5440.735 · Cited by 3,876
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Liu, J., Lam, J. W. Y., & Tang, B. Z. (2009) · Chemical Reviews · doi:10.1021/cr900149d · Cited by 1,196
Gehlen, M. H. (2020) · Journal of Photochemistry and Photobiology C: Photochemistry Reviews · doi:10.1016/j.jphotochemrev.2019.100338 · Cited by 734
Enzyme-responsive polymeric assemblies, nanoparticles and hydrogels
Hu, J., Zhang, G., & Liu, S. (2012) · Chemical Society Reviews · doi:10.1039/c2cs35103j · Cited by 722
Feng, X., Liu, L., Wang, S., & Zhu, D. (2010) · Chemical Society Reviews · doi:10.1039/b909065g · Cited by 621
High-throughput screening assays for the identification of chemical probes
Inglese, J., Johnson, R. L., Simeonov, A., Xia, M., Zheng, W., Austin, C. P., & Auld, D. S. (2007) · Nature Chemical Biology · doi:10.1038/nchembio.2007.17 · Cited by 614
Metadata
- Authors
- Kumaraswamy, S., Bergstedt, T., Shi, X., Rininsland, F., Kushon, S., Xia, W., Ley, K., Achyuthan, K., McBranch, D., & Whitten, D.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 2004-05-10
- Volume
- 101
- Issue
- 20
- Pages
- 7511-7515
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W1972158411
- Cited by
- 216
- References
- 2
- FWCI
- 7.42
- Type
- journal article
- Language
- en
- Open access
- green
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- Kumaraswamy, S., Bergstedt, T., Shi, X., Rininsland, F., Kushon, S., Xia, W., Ley, K., Achyuthan, K., McBranch, D., & Whitten, D. (2004). Fluorescent-conjugated polymer superquenching facilitates highly sensitive detection of proteases. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.0402367101
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