THE AMILORIDE-SENSITIVE NA+/H+ ANTIPORT IN 3T3-FIBROBLASTS - CHARACTERIZATION AND STIMULATION BY SERUM

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  • 1 January 1983
    • journal article
    • research article
    • Vol. 258  (10) , 6272-6276
Abstract
BALB/c 3T3 [mouse] fibroblasts have an amiloride-sensitive Na+ uptake mechanism which is hardly detectable under normal physiological conditions. The activity of this Na+ transport system can be increased to a large extent by treatments that decrease the internal pH such as loss of intracellular NH4+ as NH3 or incubation with nigericin in the presence of a low external K+ concentration. These treatments made possible an analysis of the interaction of the Na+/H+ antiport with amiloride and of the external pH dependence of the system. The addition of fetal bovine serum to quiescent 3T3 cells stimulates the initial rate of the amiloride-sensitive 22Na+ uptake by only 50%. However, after treatment of the cells with ammonia or nigericin, serum produces a 40-fold stimulation of the rate of the amiloride-sensitive 22Na+ uptake. Control experiments show that serum does not stimulate the activity of the Na+/H+ antiport by an indirect mechanism involving a depolarization of the membrane or a modification of the internal Ca2+ concentration. Some serum component may directly interact with the Na+/H+ exchanger to modify its catalytic properties.