Referenced in: none

Automatically associated channels: Kv2.1

Title: Measurement of Na+-K+ coupling ratio of Na+-K+-ATPase in rabbit proximal tubules.

Authors: M J Avison, S R Gullans, T Ogino, G Giebisch, R G Shulman

Journal, date & volume: Am. J. Physiol., 1987 Jul , 253, C126-36

PubMed link: http://www.ncbi.nlm.nih.gov/pubmed/2440310

Abstract
A combination of 23Na nuclear magnetic resonance (NMR) spectroscopy and a K+-selective electrode was used to make simultaneous measurements of net Na+ and K+ fluxes across plasma membranes of rabbit renal proximal tubules after an abrupt stimulation of Na+-K+-ATPase. After a step in extracellular K+ concentration ([K+]o) from low to higher concentration (0.1-0.3 mM to 0.5-5.2 mM) at 25 degrees C, net extrusion of Na+ and uptake of K+ were observed. These fluxes were completely inhibited by ouabain (10(-3) M). Because initial rates of K+ uptake in presence or absence of Ba2+ (a known inhibitor of plasma membrane K+ conductance) were indistinguishable, net K+ flux was virtually unidirectional. Because suspension buffers contained neither glucose nor amino acids and the ratio of net Na+ and K+ fluxes (JNa and JK, respectively) was constant over a wide range of transmembrane Na+ gradients and absolute values of the JNa and JK, it is likely that changes in electrogenic or passive net fluxes across plasma membranes were insignificant in the first 30-45 s after the [K+]o step. Thus the ratio of these initial net Na+ and K+ fluxes corresponds closely to the Na+-K+ coupling ratio of the Na+-K+-ATPase. In 12 experiments, the measured Na+-K+-ATPase coupling ratio was 1.54 +/- 0.07 (SE). The coupling ratio was constant over a wide range of intracellular Na+ content, intracellular sodium concentration, [K+]o and transmembrane Na+ gradient. The coupling ratio also remained constant over an eightfold range of Na+-K+-ATPase rates.