Abstract
Small molecules acting as selective activators (potentiators), inhibitors, or “correctors” of the CFTR chloride channel represent candidate drugs for various pathological conditions including cystic fibrosis and secretory diarrhea. The identification of CFTR pharmacological modulators may be achieved by screening highly diverse synthetic or natural compound libraries using high-throughput methods. A convenient assay for CFTR function is based on the halide sensitivity of the yellow fluorescent protein (YFP). CFTR activity can be simply assessed by measuring the rate of YFP signal decrease caused by iodide influx. This assay can be automated to test thousands of compounds per day.
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References
Verkman, A. S., and Galietta, L. J. (2009) Chloride channels as drug targets. Nat. Rev. Drug Discov. 8, 153–171.
Jayaraman, S., Haggie, P., Wachter, R. M., Remington, S. J., and Verkman, A. S. (2000) Mechanism and cellular applications of a green fluorescent protein-based halide sensor. J. Biol. Chem. 275, 6047–6050.
Galietta, L. J., Haggie, P. M., and Verkman, A. S. (2001) Green fluorescent protein-based halide indicators with improved chloride and iodide affinities. FEBS Lett. 499, 220–224.
Ma, T., Thiagarajah, J. R., Yang, H., Sonawane, N. D., Folli, C., Galietta, L. J., et al. (2002) Thiazolidinone CFTR inhibitor identified by high-throughput screening blocks cholera toxin-induced intestinal fluid secretion. J. Clin. Invest. 110, 1651–1658.
Yang, H., Shelat, A. A., Guy, R. K., Gopinath, V. S., Ma, T., Du, K., et al. (2003) Nanomolar affinity small molecule correctors of defective Delta F508-CFTR chloride channel gating. J. Biol. Chem. 278, 35079–35085.
Pedemonte, N., Sonawane, N. D., Taddei, A., Hu, J., Zegarra-Moran, O., Suen, Y. F. et al. (2005) Phenylglycine and sulfonamide correctors of defective deltaF508 and G551D cystic fibrosis transmembrane conductance regulator chloride-channel gating. Mol. Pharmacol. 67, 1797–1807.
Pedemonte, N., Lukacs, G. L., Du, K., Caci, E., Zegarra-Moran, O., Galietta, L. J. et al. (2005) Small-molecule correctors of defective DeltaF508-CFTR cellular processing identified by high-throughput screening. J. Clin. Invest. 115, 2564–2571.
Pedemonte, N., Caci, E., Sondo, E., Caputo, A., Rhoden, K., Pfeffer, U. et al. (2007) Thiocyanate transport in resting and IL-4-stimulated human bronchial epithelial cells: Role of pendrin and anion channels. J. Immunol. 178, 5144–5153.
Caputo, A., Caci, E., Ferrera, L., Pedemonte, N., Barsanti, C., Sondo, E. et al. (2008) TMEM16A, a membrane protein associated with calcium-dependent chloride channel activity. Science. 322, 590–594.
Caci, E., Caputo, A., Hinzpeter, A., Arous, N., Fanen, P., Sonawane, N. et al. (2008) Evidence for direct CFTR inhibition by CFTR(inh)-172 based on Arg347 mutagenesis. Biochem. J. 413, 135–142.
Caputo, A., Hinzpeter, A., Caci, E., Pedemonte, N., Arous, N., Di Duca, M. et al. (2009) Mutation-specific potency and efficacy of cystic fibrosis transmembrane conductance regulator chloride channel potentiators. J. Pharmacol. Exp. Ther. 330, 783–791.
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Pedemonte, N., Zegarra-Moran, O., Galietta, L.J. (2011). High-Throughput Screening of Libraries of Compounds to Identify CFTR Modulators. In: Amaral, M., Kunzelmann, K. (eds) Cystic Fibrosis. Methods in Molecular Biology, vol 741. Humana Press. https://doi.org/10.1007/978-1-61779-117-8_2
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DOI: https://doi.org/10.1007/978-1-61779-117-8_2
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