Substrate specificity of the mammary tissue anionic amino acid carrier operating in the cotransport and exchange modes

Abstract

The substrate specificity of the rat mammary tissue high affinity, Na +-dependent anionic amino acid transport system has been investigated using explants and the perfused mammary gland. d-Aspartate appears to be transported via the high affinity, Na + -dependent l-glutamate carrier. Thus, d-aspartate transport by rat mammary tissue was Na +-dependent and saturable with respect to extracellular d-aspartate with a K m and V max of 32.4 μM and 49.0 nmol/2 min per g of cells respectively. The uptake of d-aspartate by mammary explants was cis-inhibited by l-glutamate and l-aspartate, but not by d-glutamate. l-glutamate uptake by mammary tissue explants was cis-inhibited by β-glutamate, l-cysteate, l-cysteine sulfinate and dihydrokainate but not by dl- α-aminoadipate. In addition, dihydrokainate, but not dl- α-aminoadipate inhibited d-aspartate and l-glutamate uptake by the perfused gland. d-Aspartate efflux from mammary tissue explants was trans-accelerated by external l-glutamate in a dose-dependent fashion (50-500 μM). The effect of l-glutamate on d-aspartate efflux was dependent on the presence of extracellular Na +. d-Aspartate, l-aspartate and l-cysteine sulfinate (at 500 μM) also markedly trans-stimulated d-aspartate efflux from mammary tissue explants. In contrast, l-cysteine, d-glutamate, l-leucine, dihydrokainate and dl- α-aminoadipate were either weak stimulators of d-aspartate efflux or were without effect. d-Aspartate efflux from the perfused mammary gland was trans-stimulated by l-glutamate but not by d-glutamate and only weakly by l-cysteine (all at 500 μM). It appears that the mammary tissue high affinity anionic amino acid carrier can operate in the exchange mode with a similar substrate specificity to that of the co-transport mode.