Transport of potassium in Chara australis: II. Kinetics of a symport with sodium.

An electrogenic K(+)-Na+ symport with a high affinity for K+ has been found in Chara (Smith & Walker, 1989). Under voltage-clamp conditions, the symport shows up as a change in membrane current upon adding either K+ or Na+ to the bathing medium in the presence of the other. Estimation of kinetic parameters for this transport has been difficult when using intact cells, since K(+)-Na+ current changes show a rapid falling off with time at K+ concentrations above 50 microM. Cytoplasm-enriched cell fragments are used to overcome this difficulty, since they do not show the rapid falling off of current change seen with intact cells. Current-voltage curves for the membrane in the absence or presence of either K+ or Na+ are obtained, yielding difference current-voltage curves which isolate the symport currents from other transport processes. The kinetic parameters describing this transport are found to be voltage dependent, with Km for K+ ranging from 30 down to 2 microM as membrane potential varies from -140 to -400 mV, and Km for Na+ ranging between 470 and 700 microM over a membrane potential range of -140 to -310 mV. Two different models for this transport system have been investigated. One of these involves the simultaneous transport of both the driver and substrate ions across the membrane, while the other allows for the possibility of the two ions being transported consecutively in two distinct reaction steps. The experimental results are shown to be consistent with either of these cotransport models, but they do suggest that binding of K+ occurs before that of Na+, and that movement of charge across the membrane (the voltage-dependent step) occurs when the transport protein has neither K+ nor Na+ bound to it.