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J. Biol. Chem., Vol. 263, Issue 36, 19494-19497, Dec, 1988

Mechanism of cis- and trans-substrate interactions at the tetraethylammonium/H+ exchanger of rabbit renal brush-border membrane vesicles

SH Wright and TM Wunz
Department of Physiology, College of Medicine, University of Arizona, Tucson 85724.

The kinetic basis for trans-effects of intravesicular substrates on the uptake of the organic cation, tetraethylammonium (TEA), into rabbit renal brush-border membrane vesicles (BBMV) was studied. Preloading BBMV with 1, 2, or 4 mM TEA stimulated the initial rate of uptake and the total net accumulation of 0.1 mM [3H]TEA. The stimulatory effect of intravesicular TEA on the initial rate of uptake was a saturable function of the trans-TEA concentration, with a half-maximal effect noted at an intravesicular concentration of 0.28 mM. A 1 mM trans- concentration of TEA increased the Jmax of [3H]TEA uptake (from 4.3 to 6.8 nmol.mg-1.min-1) without affecting the apparent Kt. An outwardly directed H+ gradient also increased Jmax (to 10.7 nmol.mg-1.min-1), although the addition of an outwardly directed TEA gradient did not produce further increases in the rate of TEA uptake. External H+ acted as a competitive inhibitor of TEA uptake, and an increase in external [H+] (from 32 nM to 100 nM) produced an increase in the apparent Kt for TEA transport (from 0.12 to 0.26 mM) without affecting the Jmax. The results suggested that TEA and H+ compete for a common site or set of mutually exclusive sites on the cytoplasmic and luminal aspects of TEA/H+ exchanger in the renal brush border, and that these sites have a similar affinity for TEA.
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