1. In isolated hepatocytes NaF increased the rate of 45Ca2+ exchange, the cytoplasmic free Ca2+ concentration ([Ca2+]i) (monitored by using quin2), and the activity of glycogen phosphorylase a in a Ca2+-dependent manner. 2. In cells previously incubated in the absence of extracellular Ca2+(Ca2+o), NaF caused a pronounced enhancement in the increases in the activity of glycogen phosphorylase and in [Ca2+]i observed when Ca2+ was subsequently added. The effect of NaF on glycogen phosphorylase activity was inhibited by verapamil and deferoxamine, and was potentiated by AlCl3. 3. The actions of NaF were associated with (a) increases in [3H]inositol polyphosphates, which were slower in onset and about half the magnitude of those induced by vasopressin, in hepatocytes labelled with [3H]inositol, and (b) enhanced rates of O2 utilization and decreased concentrations of ATP. The latter effects were not potentiated by AlCl3. 4. Preincubation of hepatocytes with vasopressin in the absence of added Ca2+o for times up to 30 min did not diminish the ability of a subsequent addition of extracellular Ca2+ to activate glycogen phosphorylase. 5. 12-O-Tetradecanoylphorbol 13-acetate had little effect on 45Ca2+ exchange and did not enhance the activation by Ca2+o of phosphorylase in hepatocytes incubated in the absence of Ca2+o. 6. On the basis of the observation that AlF4- activates GTP-binding regulatory proteins [Sternweiss & Gilman (1982) Proc. Natl. Acad. Sci. U.S.A. 79, 4888-4891], it is concluded that the present results provide evidence for the function of a GTP-binding regulatory protein in the mechanism by which hormones stimulate plasma-membrane Ca2+ inflow in the liver cell, and indicate that an increase in [Ca2+]i and the activation of protein kinase C are not part of this mechanism.
The stimulation by sodium fluoride of plasma-membrane Ca2+ inflow in isolated hepatocytes. Evidence that a GTP-binding regulatory protein is involved in the hormonal stimulation of Ca2+ inflow
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B P Hughes, G J Barritt; The stimulation by sodium fluoride of plasma-membrane Ca2+ inflow in isolated hepatocytes. Evidence that a GTP-binding regulatory protein is involved in the hormonal stimulation of Ca2+ inflow. Biochem J 1 July 1987; 245 (1): 41–47. doi: https://doi.org/10.1042/bj2450041
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