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Title: Pre-steady-state and reverse transport currents in the GABA transporter GAT1. Author: Cherubino F, Bertram S, Bossi E, Peres A. Journal: Am J Physiol Cell Physiol; 2012 Apr 15; 302(8):C1096-108. PubMed ID: 22173867. Abstract: The role of internal substrates in the biophysical properties of the GABA transporter GAT1 has been investigated electrophysiologically in Xenopus oocytes heterologously expressing the cotransporter. Increments in Cl(-) and/or Na(+) concentrations caused by intracellular injections did not produce significant effects on the pre-steady-state currents, while a positive shift of the charge-voltage (Q-V) and decay time constant (τ)-voltage (τ-V) curves, together with a slowing of τ at positive potentials, was observed following treatments producing cytosolic Cl(-) depletion. Activation of the reverse transport mode by injections of GABA caused a reduction in the displaced charge. In the absence of external Cl(-), a stronger reduction in the displaced charge, together with a significant increase in reverse transport current, was observed. Therefore, complementarity between pre-steady-state and transport currents, observed in the forward mode, is preserved in the reverse mode. All these findings can be qualitatively reproduced by a kinetic scheme in which, in the forward mode, the Cl(-) ion is released first, after the inward charge movement, while the two Na(+) ions can be released only after binding of external GABA. In the reverse mode, internal GABA must bind first to the empty transporter, followed by internal Na(+) and Cl(-).[Abstract] [Full Text] [Related] [New Search]