The glutamine/amino acid transporter (ASCT2) reconstituted in liposomes: electrical nature of the glutamine/glutamate antiport.

Abstract

The glutamine/amino acid transporter solubilized from rat renal apical plasma membrane (brush-border membrane) with C12E8 and reconstituted into liposomes has been previously identified as the ASCT2 transporter. The reconstituted transporter catalyses an antiport reaction in which extraliposomal glutamine and Na+ are cotransported in exchange with intraliposomal neutral amino acids. Differently from other neutral amino acid transporters, ASCT2 accepts also glutamate as substrate, as demonstrated by the glutamine/glutamate antiport measured in proteoliposomes. The electrical nature of the homologous glutamine/glutamine antiport and of the heterologous glutamine/glutamate antiport has been investigated by imposing a K+ diffusion potential (positive outside) across the proteoliposomal membrane in the presence of valinomycin. The membrane potential did not affect the glutamine/glutamine antiport whereas it stimulated about two fold the glutamine/glutamate antiport rate.