Abstract
The transport of three Na+ per two K+ means that the Na,K-ATPase is electrogenic, and though the currents generated by the ion pump are small compared to ion channel currents, they can be measured using electrophysiology, both steady-state pumping and individual steps in the transport cycle. Various electrophysiological techniques have been used to study the endogenous pumps of the squid giant axon and of cardiac myocytes from for example rabbits. Here, we describe the characterization of heterologously expressed Na,K-ATPases using two-electrode voltage clamping (TEVC) and oocytes from the Xenopus laevis frog as the model cell. With this system, the effects of particular mutations can be studied, including the numerous mutations that in later years have been found to cause human diseases.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Skou JC (1957) The influence of some cations on an adenosine triphosphatase from peripheral nerves. J Am Soc Nephrol 23:394–401
Albers RW, Fahn S, Koval GJ (1963) The role of sodium ions in the activation of electrophorus electric organ adenosine triphosphatase. Proc Natl Acad Sci U S A 50(3):474–481
Glitsch HG (2001) Electrophysiology of the sodium-potassium-ATPase in cardiac cells. Physiol Rev 81(4):1791–1826
Holmgren M et al (2000) Three distinct and sequential steps in the release of sodium ions by the Na+/K+-ATPase. Nature 403(6772):898–901
Noguchi S, Mishina M, Kawamura M, Numa S (1987) Expression of functional (Na+ + K+)-ATPase from cloned cDNAs. FEBS Lett 225(1–2):27–32
Poulsen H et al (2010) Neurological disease mutations compromise a C-terminal ion pathway in the Na(+)/K(+)-ATPase. Nature 467(7311):99–102
Azizan EA et al (2013) Somatic mutations in ATP1A1 and CACNA1D underlie a common subtype of adrenal hypertension. Nat Genet 45(9):1055–1060
Li M et al (2015) A functional correlate of severity in alternating hemiplegia of childhood. Neurobiol Dis 77:88–93
Price EM, Lingrel JB (1988) Structure-function relationships in the Na, K-ATPase alpha subunit: site-directed mutagenesis of glutamine-111 to arginine and asparagine-122 to aspartic acid generates a ouabain-resistant enzyme. Biochemistry 27(22):8400–8408
Li C, Geering K, Horisberger JD (2006) The third sodium binding site of Na, K-ATPase is functionally linked to acidic pH-activated inward current. J Membr Biol 213(1):1–9
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
Hilbers, F., Poulsen, H. (2016). Electrophysiological Characterization of Na,K-ATPases Expressed in Xenopus laevis Oocytes Using Two-Electrode Voltage Clamping. In: Bublitz, M. (eds) P-Type ATPases. Methods in Molecular Biology, vol 1377. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3179-8_27
Download citation
DOI: https://doi.org/10.1007/978-1-4939-3179-8_27
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3178-1
Online ISBN: 978-1-4939-3179-8
eBook Packages: Springer Protocols