G. Barmashenko et al., Depolarization increases the apparent affinity of the Na+-K+ pump to cytoplasmic Na+ in isolated guinea-pig ventricular myocytes, J PHYSL LON, 517(3), 1999, pp. 691-698
1. In order to investigate the possible effect of membrane potential on cyt
oplasmic Na+ binding to the Na+-K+ pump, we studied Na+-K+ pump current-vol
tage relationships in single guinea-pig ventricular myocytes whole-cell vol
tage clamped with pipette solutions containing various concentrations of Na
+ ([Na+](pip)) and either tetraethylammonium (TEA(+)) or N-methyl-D-glucami
ne (NMDG(+)) as the main cation. The experiments were conducted at 30 degre
es C under conditions designed to abolish the known voltage dependence of o
ther steps in the pump cycle, i.e. in Na+-free external media containing 20
mM Cs+.
2. Na+-K+ pump current (I-p) was absent in cells dialysed with Na+-free pip
ette solutions and was almost voltage independent at 50 mM Na-pip(+) (poten
tial range: -100 to +40 mV). By contrast, the activation of I-p by 0.5-5 mM
Na-pip(+) was clearly voltage sensitive and increased with depolarization,
independently of the main intracellular cation species.
3. The apparent affinity of the Na+-K+ pump for cytoplasmic Na+ increased m
onotonically with depolarization. The [Na+](pip) required for half-maximal
I-p activation (K-0.5 value) amounted to 5.6 mar at -100 mV and to 2 2 mM a
t +40 mV.
4. The results suggest that cytoplasmic Na+ binding and/or a subsequent par
tial reaction in the pump cycle prior to Na+ release is voltage dependent.
From the voltage dependence of the K-0.5 values the dielectric coefficient
for intracellular Na+ binding/translocation was calculated to be similar to
0.08. The voltage-dependent mechanism might add to the activation of the c
ardiac Na+-K+ pump during cardiac excitation.