Abstract
Cystic fibrosis transmembrane conductance regulator (CFTR)-mediated chloride secretion is critical to maintaining airway surface hydration and efficient mucociliary clearance in the upper airways. Mutations in CFTR in cystic fibrosis lead to reduced expression of functional CFTR channels at the apical plasma membrane of the airway epithelium, leading to dehydration of the airway surface liquid and diminished mucociliary clearance. Cell surface CFTR is modulated by changes in CFTR endocytosis and recycling, effectively altering the cell surface abundance of the channel. This chapter examines current methods employed to measure the cell surface expression of CFTR, as well as methods to monitor CFTR movement through the endocytic pathway.
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References
Guggino, W. B., and Stanton, B. A. (2006) New insights into cystic fibrosis: Molecular switches that regulate CFTR. Nat. Rev. Mol. Cell. Biol. 7, 426–436.
Gruenberg, J., and van der Goot, F. G. (2006) Mechanisms of pathogen entry through the endosomal compartments. Nat. Rev. Mol. Cell. Biol. 7, 495–504.
Swiatecka-Urban, A., Boyd, C., Coutermarsh, B., Karlson, K. H., Barnaby, R., Aschenbrenner, L., et al. (2004) Myosin VI regulates endocytosis of the cystic fibrosis transmembrane conductance regulator. J. Biol. Chem. 279, 38025–38031.
Bomberger, J. M., Barnaby, R. L., and Stanton, B. A. (2009) The deubiquitinating enzyme USP10 regulates the post-endocytic sorting of cystic fibrosis transmembrane conductance regulator in airway epithelial cells. J. Biol. Chem. 284, 18778–18789.
Swiatecka-Urban, A., Talebian, L., Kanno, E., Moreau-Marquis, S., Coutermarsh, B., Hansen, K., et al. (2007) Myosin VB is required for trafficking of CFTR in RAB11A-specific apical recycling endosomes in polarized human airway epithelial cells. J. Biol. Chem. 282, 23725–23736.
Biemesderfer, D., Mentone, S. A., Mooseker, M., and Hasson, T. (2002) Expression of myosin VI within the early endocytic pathway in adult and developing proximal tubules. Am. J. Physiol. Renal Physiol. 282, F785–F794.
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Bomberger, J.M., Guggino, W.B., Stanton, B.A. (2011). Methods to Monitor Cell Surface Expression and Endocytic Trafficking of CFTR in Polarized Epithelial Cells. 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_18
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DOI: https://doi.org/10.1007/978-1-61779-117-8_18
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Online ISBN: 978-1-61779-117-8
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