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Biophys J, July 2000, p. 215-230, Vol. 79, No. 1
Institute of Physiology, University of Zurich, Zurich, Switzerland
In the kidney proximal tubule, acidification of the
glomerular filtrate leads to an inhibition of inorganic phosphate
(Pi) reabsorption by type II Na+-coupled
cotransporters (NaPi-II). As external pH also alters the
divalent/monovalent Pi ratio, it has been difficult to
separate putative proton interactions with the cotransporter from
direct titration of divalent Pi, the preferred species
transported. To distinguish between these possibilities and identify
pH-sensitive transitions in the cotransport cycle, the pH-dependent
kinetics of two NaPi-II isoforms, expressed in Xenopus
laevis oocytes, were investigated
electrophysiologically. At
50 mV, both isoforms showed >70%
suppression of electrogenic response for an external pH change from 8.0 to 6.2, not attributable to titration of divalent Pi. This
was accompanied by a progressive removal of steady-state voltage
dependence. The NaPi-II-related uncoupled slippage current was
unaffected by a pH change from 7.4 to 6.2, with no shift in the
reversal potential, which suggested that protons do not function as
substrate. The voltage-dependence of pre-steady-state relaxations was
shifted to depolarizing potentials in 100 mM and 0 mM
Naext+ and two kinetic components were resolved, the
slower of which was pH-dependent. The changes in kinetics are predicted
by a model in which protons interact with the empty carrier and final
Na+ binding step.
Biophys J, July 2000, p. 215-230, Vol. 79, No. 1
© 2000 by the Biophysical Society 0006-3495/00/07/215/16 $2.00
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