Chemiluminescence Assays to Investigate Membrane Expression and Clathrin-Mediated Endocytosis of KATP Channels

  • Andrew J. Smith
  • Asipu Sivaprasadarao
Part of the Methods in Molecular Biology book series (MIMB, volume 491)


Macroscopic ion channel currents (I) are a product of the channel open probability (P o), the single channel current (i) and the number of channels present on the cell surface (N) at any given time (I = P o iN). Endocytosis has been shown to be one of the key determinants of cell surface channel density and the defects of this process have been linked to diseases relating to ion channel dysfunction. Chemiluminescence-based techniques provide a rapid method for the examination of the rates of endocytosis and steady-state cell surface density of ion channels and have previously been used to investigate the endocytosis of pancreatic ATP-sensitive potassium (KATP) channels.

Key words

Endocytosis Internalisation Chemiluminescence Surface density KATP channels. 



The authors would like to thank Dr. Jamel Mankouri for the development of the chemiluminescence-based assays.


  1. 1.
    Hu, K., Huang, C. S., Jan, Y. N., and Jan, L. Y. (2003) ATP-sensitive potassium channel traffic regulation by adenosine and protein kinase C. Neuron 38, 417–32.CrossRefPubMedGoogle Scholar
  2. 2.
    Okiyoneda, T. and Lukacs, G. L. (2007) Cell surface dynamics of CFTR: the ins and outs. Biochim Biophys Acta 1773, 476–9.CrossRefPubMedGoogle Scholar
  3. 3.
    Herfst, L. J., Rook, M. B., and Jongsma, H. J. (2004) Trafficking and functional expression of cardiac Na+ channels. J Mol Cell Cardiol 36, 185–93.CrossRefPubMedGoogle Scholar
  4. 4.
    Steele, D. F., Eldstrom, J., and Fedida, D. (2007) Mechanisms of cardiac potassium channel trafficking. J Physiol 582, 17–26.CrossRefPubMedGoogle Scholar
  5. 5.
    Jarvis, S. E. and Zamponi, G. W. (2007) Trafficking and regulation of neuronal voltage-gated calcium channels. Curr Opin Cell Biol 19, 474–82.CrossRefPubMedGoogle Scholar
  6. 6.
    Mankouri, J., Taneja, T. K., Smith, A. J., Ponnambalam, S., and Sivaprasadarao, A. (2006) Kir6.2 mutations causing neonatal diabetes prevent endocytosis of ATP-sensitive potassium channels. EMBO J 25, 4142–51.CrossRefPubMedGoogle Scholar
  7. 7.
    Zerangue, N., Schwappach, B., Jan, Y. N., and Jan, L. Y. (1999) A new ER trafficking signal regulates the subunit stoichiometry of plasma membrane K(ATP) channels. Neuron 22, 537–48.CrossRefPubMedGoogle Scholar

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Andrew J. Smith
    • 1
  • Asipu Sivaprasadarao
    • 1
  1. 1.Institute of Membrane and Systems BiologyUniversity of LeedsLeedsUK

Personalised recommendations