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J. Biol. Chem., Vol. 268, Issue 34, 25527-25535, 12, 1993
SA Levine, MH Montrose, CM Tse and M Donowitz
The kinetics and second messenger regulation of three cloned mammalian
intestinal Na+/H+ exchangers were studied using fluorometric techniques.
These exchangers, NHE1, NHE2, and NHE3, were stably expressed in PS120
fibroblasts, which lack an endogenous Na+/H+ exchanger. H+ kinetic data
indicated cooperativity by internal protons, with Hill coefficients of
approximately 2 for all three isoforms. In contrast, Na+ kinetic data fit
Michaelis-Menten kinetics, with Km (Na+) 15-18 mM and a Hill coefficient of
approximately 1. The exchangers were all activated by growth factors and
thrombin; in NHE1 these agonists increased the apparent affinity for
intracellular H+, but did not change Vmax, while for NHE2 and NHE3 the
effect was on Vmax alone. Phorbol ester stimulated NHE1 and NHE2, but
inhibited NHE3 with a decrease in Vmax. ATP-depletion decreased Vmax and
the apparent affinity for H+ for all three isoforms, and reduced the Hill
coefficient to approximately 1, suggesting that a basal level of
phosphorylation was required for the cooperativity. The differences in
kinetics and second messenger regulation suggest that the NHE isoforms may
serve different cellular functions. The up- and down-regulation of NHE3 by
kinases indicates that this isoform may be involved in a specialized
function such as Na+ absorption.
Kinetics and regulation of three cloned mammalian Na+/H+ exchangers stably expressed in a fibroblast cell line
Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205-2195.
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S. Akhter, S. K. Nath, C. M. Tse, J. Williams, M. Zasloff, and M. Donowitz Squalamine, a novel cationic steroid, specifically inhibits the brush-border Na+/H+ exchanger isoform NHE3 Am J Physiol Cell Physiol, January 1, 1999; 276(1): C136 - C144. [Abstract] [Full Text] [PDF]
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G. Lamprecht, E. J. Weinman, and C.-H. C. Yun The Role of NHERF and E3KARP in the cAMP-mediated Inhibition of NHE3 J. Biol. Chem., November 6, 1998; 273(45): 29972 - 29978. [Abstract] [Full Text] [PDF]
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