The voltage-dependent step of the chloride transporter of Valonia utricularis encounters a Nernst-Planck and not an Eyring type of potential energy barrier.
Voltage-clamp experiments were performed on cells of the giant marine alga Valonia utricularis to study the voltage dependence of the previously postulated chloride transporter (Wang, J., G. Wehner, R. Benz, and U. Zimmermann. 1991. Biophys. J. 59:235-248). Only one exponential current relaxation (apart from the capacitive spike) could be resolved up to a clamp voltage of approximately 120 mV within the time resolution of our experimental instrumentation (100 mus). This means that the rate constants of the heterogeneous complexation, k(R) (association) and k(D) (dissociation), were too fast to be resolved. Therefore, the "Läuger" model for carrier-mediated ion transport with equilibrium heterogeneous surface reaction was used to fit the experimental results. The voltage dependence of the initial membrane conductance was used for the evaluation of the voltage dependence of the translocation rate constant of the complexed carriers, k(AS). The initial conductance was found to be independent on the clamp voltage, which means that the translocation rate constant k(AS) is a linear function of the applied voltage and that the voltage dependence of the translocation of charged carriers through the plasmalemma could be explained by a square-type Nernst-Planck barrier. The movement of the complexed form of the carrier through the membrane may be explained by a diffusion process rather than by simple first-order kinetic jump across an Eyring-type potential well. The current relaxation after a voltage clamp was studied as a function of the external chloride concentration. The results allowed an estimation of the stability constant, K, of the heterogeneous complexation reaction and a calculation of the translocation rate constants of the free and the complexed carriers, k(s) and k(AS), respectively.
Lehrstuhl für Biotechnologie, Biozentrum der Universität Würzburg, D-8700 Würzburg, Germany.
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J
Wang
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U
Zimmermann
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R
Benz