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The amiloride sensitive sodium proton antiport in 3t3 fibroblasts characterization and stimulation by serum



The amiloride sensitive sodium proton antiport in 3t3 fibroblasts characterization and stimulation by serum



Journal of Biological Chemistry 258(10): 6272-6276



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.

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Accession: 006599078

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