Suppression of the plasma membrane H+-conductance on the background of high H+-pump activity in dithiothreitol-treated Chara cells
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Photosynthesizing cells of characean algae exposed to light are able to produce pH bands corresponding to alternate areas with dominant H+-pump activity and high H+-conductance of the cell membrane. The action potential generation temporally arrests the counter-directed H+ fluxes, which gives rise to opposite pH shifts in different cell regions and represents a suitable indicator for activities of the plasma membrane H+-transporting systems. Measurements of pH near the cell surface by means of microelectrodes and microspectrophotometry in the presence of pH-indicating dye thymol blue have shown that the treatment of cells with dithiothreitol (SH-group reducing agent) suppresses pH changes induced by the action potential generation in the alkaline cell areas and considerably increases the concurrent pH changes in the acid regions. Measurements of plasma membrane resistance in the alkaline zones revealed that dithiothreitol inhibits the light-dependent conductance of the resting cell and diminishes the conductance inactivation caused by the action potential generation. The data suggest that the reduction of accessible disulfide bonds results in the decrease of H+-conductance, whereas the activity of plasma membrane H+-pump remains unimpaired or is even enhanced.
KeywordsCharacean algae plasmalemma H+ transport membrane conductance dithiothreitol action potential
pH in the outer medium in the proximity to cell surface
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