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Received July 19, 2002
Accepted after revision August 30, 2002
1 Institute of Physiology, Humboldt University, Tucholskystrae 2, D 10117 Berlin, Germany
2 Institute of Physiology, Humboldt University, Tucholskystrae 2, D 10117 Berlin, Germany, and Department of Neonatology, Humboldt University, Augustenburger Platz 1, D 13353 Berlin, Germany
3 Department of Molecular Biophysics and Physiology, Rush Presbyterian St Luke's Medical Center, 1750 West Harrison Street, Chicago, IL 60612-3824, USA
4 Institute of Physiology, Humboldt University, Tucholskystrae 2, D-10117 Berlin, Germany
* To whom correspondence should be addressed. E-mail: claudia.eder{at}charite.de.
Voltage-activated proton currents are reported for the first time in human peripheral blood T and B lymphocytes and in the human leukaemic T cell line Jurkat E6-1. The properties of H+ currents studied using tight-seal voltage-clamp recording techniques were similar in all cells. Changing the pH gradient by one unit caused a 47 mV shift in the reversal potential, demonstrating high selectivity of the channels for protons. H+ current activation upon membrane depolarisation had a sigmoidal time course that could be fitted by a single exponential equation after a brief delay. Increasing pHo shifted the activation threshold to more negative potentials, and increased both the H+ current amplitude and the rate of activation. In lymphocytes studied at pHi 6.0, the activation threshold was more negative and the H+ current density was three times larger than at pHi 7.0. Increasing the intracellular Ca2+ concentration to 1 µM did not change H+ current amplitude or kinetics detectably. Extracellularly applied Zn2+ and Cd2+ inhibited proton currents, slowing activation and shifting the voltage-activation curve to more positive potentials. The H+ current amplitude was 100 times larger in CD19+ B lymphocytes and in Jurkat E6-1 cells than in CD3+ T lymphocytes. Following stimulation with the phorbol ester PMA, the H+ current density in peripheral blood T lymphocytes and Jurkat T cells increased. In contrast, the H+ current density of phorbol acetate (PMA)-stimulated B lymphocytes was reduced and activation became slower. The pattern of expression of H+ channels in lymphocytes appears well suited to their proposed role of charge compensation during the respiratory burst.
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