Partial purification of a potassium channel with low permeability for sodium from tonoplast membranes of Hordeum vulgare cv. Gerbel.
A potassium-specific tonoplast channel was identified by reconstitution of tonoplast polypeptides into planar lipid bilayer membranes. Highly purified tonoplast membranes were solubilized in Triton X-100-containing buffer and fractionated by size-exclusion chromatography. The protein fractions were assayed for ion channel activity in a planar bilayer system, and the potassium channel was routinely recovered in specific fractions corresponding to an apparent molecular mass of 80 kDa. In symmetrical electrolyte solutions of 100 mM potassium chloride, the potassium channel had a single-channel conductance of 72 pS. Substates of the channel with conductances of 17, 33 and 52 pS were frequently observed. After identification of the channel in low or high KCl, addition of sodium acetate or sodium chloride caused only insignificant conductance changes. This result suggested that the channel was not or little permeable for sodium or chloride, whereas it had similar single-channel conductance for rubidium and caesium ions as compared with potassium ions. The channel is presumably responsible for the equilibration of potassium between the vacuole and the cytosol. The role of the channel in the physiology of the barley cell under salt stress is discussed.
Klughammer
B
B
Lehrstuhl für Botanik 1, Universität, Wüzburg, Germany.
Betz
M
M
Benz
R
R
Dietz
K J
KJ
D002675:Child, Preschool; D002712:Chlorides / Q000378:metabolism; D001467:Hordeum / Q000166:cytology*; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism*; D008564:Membrane Potentials; D010539:Permeability; D011188:Potassium / Q000378:metabolism; D015221:Potassium Channels / Q000378:metabolism*; D012964:Sodium / Q000378:metabolism*
D002712:Chlorides; D015221:Potassium Channels; D012964:Sodium; D011188:Potassium