ATP,pH and Mg2+ modulate a cation current in Beta vulgaris vacuoles: A possible shunt conductance for the vacuolar H+-ATPase |
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Authors: | J M Davies D Sanders |
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Institution: | (1) Plant Laboratory, Department of Biology, University of York, P.O. Box 373, Y01 5YW York, UK |
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Abstract: | Macroscopic instantaneous and time-dependent currents have been measured in the vacuolar membrane of Beta vulgaris using a patch clamp configuration analogous to whole cell mode. At low cytosolic Ca2+ and in the absence of Mg2+, only an instantaneous current was observed. This current is carried predominantly by cations (PK PCl 7 1, pna pcl 4 1 and arginine is also conducted). The instantaneous current can be activated by ATP4– (e.g., ATP-activated mean K+ current density was –20 mA.m–2 at a membrane voltage of –20 mV) and by increasing cytosolic pH and Mg2+ (raising Mg2+ from 0 to 0.4 mm induced a mean current density increase of –7 mA.m–2 at –20 mV). Such current can be activated by simultaneous addition of putative in vivo concentrations of ATP4–/MgATP/Mg
free
2+
(in the presence of bafilomycin to inhibit the vacuolar ATPase) and further modulated by cytosolic pH. With vacuolar K+ concentration greater than that of the cytosol, activation of the instantaneous current would mediate vacuolar K+ release over the range of physiological membrane voltage. It is argued that the ATP4–-activated current, in addition to acting as a K+ mobilization pathway, could provide a counter-ion (shunt) conductance, allowing the two electrogenic H+ pumps which reside in the vacuolar membrane to acidify the vacuolar lumen.A separate time-dependent current, which was not observed at low Ca2+ concentrations (less than 500 nm) could also be elicited by addition of Mg2+ at the cytoplasmic membrane face. This current was stimulated by increasing cytoplasmic pH.The authors are grateful to the BBSRC for financial support (Grant PG87/529) and to the Royal Society (University Research Fellowship to J.M.D.). We thank C. Abbott, K. Partridge and J. Robinson for plant cultivation; A. Amtmann, A. Bertl, D. Gradmann and G. Thiel for helpful discussion. |
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Keywords: | Amino acid transport ATP regulation Beta Mg2+ pH |
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