Conduction of monovalent and divalent cations in the slow vacuolar channel.
The conduction properties of individual physiologically important cations Na+, K+, Mg2+, and Ca2+ were determined in the slowly activating (SV) channel of sugar beet vacuoles. Current-voltage relationships of the open channel were measured on excised tonoplast patches in a continuous manner by applying a +/-140 mV ramp-wave protocol. Applying KCl gradients of either direction across the patch we have determined that the relative Cl- to K+ permeability was < or =1%. Symmetrical increase of the concentration of tested cation caused an increase of the single channel conductance followed by saturation. Fitting of binding isotherms at zero voltage to the Michaelis-Menten equation resulted in values of maximal conductance of 300, 385, 18, and 13 pS, and of apparent dissociation constants of 64, 103, 0.04, and 0.08 mm for Na+, K+, Mg2+, and Ca2+, respectively. Deviations from the single-ion occupancy mechanism are documented, and alternative models of permeation are discussed. The magnitude of currents carried by divalent cations at low concentrations can be explained by an unrealistically wide (approximately 140 A) radius of the pore entrance. We propose instead a fixed negative charge in the pore vestibules, which concentrates the cations in their proximity. The conduction properties of the SV channel are compared with reported characteristics of voltage-dependent Ca2+-permeable channels, and consequences for a possible reduction of postulated multiplicity of Ca2+ pathways across the tonoplast are drawn.
Pottosin
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Centro Universitario de Investigaciones Biomedicas, Av. 25 de Julio s/n, Apdo Postal/P.O. Box #199, 28047 Colima, Mexico.
Dobrovinskaya
O R
OR
Muñiz
J
J
D002118:Calcium / Q000378:metabolism / Q000494:pharmacology; D015220:Calcium Channels / Q000378:metabolism; D002413:Cations, Divalent / Q000378:metabolism*; D002414:Cations, Monovalent / Q000378:metabolism*; D002463:Cell Membrane Permeability; D019663:Chenopodiaceae / Q000378:metabolism*; D002712:Chlorides / Q000378:metabolism; D015640:Ion Channel Gating; D007473:Ion Channels / Q000378:metabolism*; D017136:Ion Transport; D007700:Kinetics; D008274:Magnesium / Q000378:metabolism / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects; D018408:Patch-Clamp Techniques; D010940:Plant Proteins / Q000378:metabolism*; D018517:Plant Roots / Q000378:metabolism; D011188:Potassium / Q000378:metabolism / Q000494:pharmacology; D012964:Sodium / Q000378:metabolism / Q000494:pharmacology; D014617:Vacuoles / Q000378:metabolism*
D015220:Calcium Channels; D002413:Cations, Divalent; D002414:Cations, Monovalent; D002712:Chlorides; D007473:Ion Channels; D010940:Plant Proteins; D012964:Sodium; D008274:Magnesium; D011188:Potassium; D002118:Calcium