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Abstract

Serotonin transporter (SERT) is the main target for widely used antidepressant agents. Several of these drugs, including imipramine, citalopram, sertraline, and fluoxetine (Prozac), bound more avidly to SERT in the presence of Cl⁻. In contrast, Cl⁻ did not enhance cocaine or paroxetine binding. A Cl⁻ binding site recently identified in SERT, and shown to be important for Cl⁻ dependent transport, was also critical for the Cl⁻ dependence of antidepressant affinity. Mutation of the residues contributing to this site eliminated the Cl⁻-mediated affinity increase for imipramine and fluoxetine. Analysis of ligand docking to a single state of SERT indicated only small differences in the energy of interaction between bound ligands and Cl⁻. These differences in interaction energy cannot account for the affinity differences observed for Cl⁻ dependence. However, fluoxetine binding led to a conformational change, detected by cysteine accessibility experiments, that was qualitatively different from that induced by cocaine or other ligands. Given the known Cl⁻ requirement for serotonin-induced conformational changes, we propose that Cl⁻ binding facilitates conformational changes required for optimal binding of fluoxetine and other antidepressant drugs.