Ionic selectivity of the coupled and uncoupled currents carried by the amino acid transporter KAAT1

The ability of the intestinal amino acid cotransporter KAAT-1 expressed in Xenopus oocytes to transport different cations in either amino acid coupled or uncoupled manner was studied using voltage-clamp conditions. KAAT1-expressing oocytes exhibit a transporter-related current in the absence of organic substrate (uncoupled current). In the presence of various alkali cations the amplitude of this current follows the sequence: I-Li>I-Na>I(K)congruent to I(Rb)congruent to I-Cs. Addition of 1 mM leucine causes large increases in K+ and Na+ currents, while the Li+ current undergoes a more complex change and Rb+ and Cs+ currents are only marginally affected. Presteady-state currents in the absence of organic substrate are apparent when Na+, K+, or Li+ are the bathing ions; analysis of these currents in terms of charge movement reveals that Na+, K+, and Li+ interact differently with the transporter The uncoupled current in mixtures of Na+ and Li+ fails to exhibit anomalous mole-fraction behavior. Kinetic analysis of ion binding and uncoupled permeation argues against a multi-ion single-file mechanism in the KAAT1 cotransporter.