Summary
The regulation of voltage-dependent Ca2+ channels by protein phosphorylation and dephosphorylation was studied using tonoplast-free cells ofNitellopsis. Since the Ca2+-channel activation has a dominant role in the membrane excitation of tonoplast-free cells (T. Shiina and M. Tazawa,J. Membrane Biol. 96:263–276, 1987), it seems to be reasonable to assume that any change of the membrane excitability reflects a modulation of the Ca2+ channel. When agents that enhance phosphoprotein dephosphorylation (protein kinase, inhibitor, phosphoprotein phosphatase-1, -2A) were introduced to the intracellular surface of the plasmalemma (twice-perfused tonoplast-free cells), the membrane potential depolarized and the membrane resistance decreased under current-clamp experiments. By contrast, when cells were challenged with agents that enhance protein phosphorylation (phosphoprotein phosphatase inhibitor-1, α-naphthylphosphate), the membrane potential hyperpolarized, and the membrane resistance increased. When phosphoprotein phosphatase-1 or -2A was perfused, the current-voltage (I–V) curve which was obtained under ramp voltage-clamp condition exhibited the so-called N-shaped characteristic, indicating an acceleration of the Ca2+-channel activation. This effect was suppressed by the addition of phosphoprotein phosphatase inhibitors. ATP-γ-S, which is assumed to stimulate protein phosphorylation, decreased the inward current in theI–V curve. The dependence of the Ca2+-channel activation on intracellular ATP was different between the once-perfused and twice-perfused cells. In once-perfused cells, the membrane excitability was reduced by low intracellular ATP concentration. By contrast, in twice-perfused cells, excitability was enhanced by ATP.
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Shiina, T., Wayne, R., Lim Tung, H.Y. et al. Possible involvement of protein phosphorylation/dephosphorylation in the modulation of Ca2+ channel in tonoplast-free cells ofNitellopsis . J. Membrain Biol. 102, 255–264 (1988). https://doi.org/10.1007/BF01925719
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DOI: https://doi.org/10.1007/BF01925719