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• W2334836183 startingPage "C1661" @default.
• W2334836183 abstract "Li + interacts with the Na + /Cl − -dependent GABA transporter, GAT1, under two conditions: in the absence of Na + it induces a voltage-dependent leak current; in the presence of Na + and GABA, Li + stimulates GABA-induced steady-state currents. The amino acids directly involved in the interaction with the Na + and Li + ions at the so-called “ Na2” binding site have been identified, but how Li + affects the kinetics of GABA cotransport has not been fully explored. We expressed GAT1 in Xenopus oocytes and applied the two-electrode voltage clamp and 22 Na uptake assays to determine coupling ratios and steady-state and presteady-state kinetics under experimental conditions in which extracellular Na + was partially substituted by Li + . Three novel findings are: 1) Li + reduced the coupling ratio between Na + and net charge translocated during GABA cotransport; 2) Li + increased the apparent Na + affinity without changing its voltage dependence; 3) Li + altered the voltage dependence of presteady-state relaxations in the absence of GABA. We propose an ordered binding scheme for cotransport in which either a Na + or Li + ion can bind at the putative first cation binding site ( Na2). This is followed by the cooperative binding of the second Na + ion at the second cation binding site ( Na1) and then binding of GABA. With Li + bound to Na2, the second Na + ion binds more readily GAT1, and despite a lower apparent GABA affinity, the translocation rate of the fully loaded carrier is not reduced. Numerical simulations using a nonrapid equilibrium model fully recapitulated our experimental findings." @default.
• W2334836183 created "2016-06-24" @default.
• W2334836183 creator A5009064853 @default.
• W2334836183 creator A5027791484 @default.
• W2334836183 date "2012-06-01" @default.
• W2334836183 modified "2023-10-18" @default.
• W2334836183 title "Using lithium to probe sequential cation interactions with GAT1" @default.
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• W2334836183 doi "https://doi.org/10.1152/ajpcell.00446.2011" @default.
• W2334836183 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/22460712" @default.
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