Serotonin transporter mutations associated with obsessive-compulsive disorder and phosphorylation alter binding affinity for inhibitors

Mutants of serotonin transporter that are altered in their regulation by cGMP were tested for the ability of cocaine and the antidepressant drugs imipramine, sertraline, citalopram and fluoxetine to inhibit serotonin transport. Mutation at Ile-425 to valine, found in some patients with obsessive-compulsive disorder, altered the response of SERT to cGMP (Kilic, F., Murphy, D.L., Rudnick, G., 2003. A human serotonin transporter mutation causes constitutive activation of transport activity. Mol. Pharmacol. 64, 440-446). This mutation selectively decreased the potency of sertraline for inhibiting serotonin transport. The potencies of imipramine, citalopram, fluoxetine and cocaine for inhibiting transport were not affected by this mutation. In binding measurements with the cocaine analog 2β-carbomethoxy-3β-(4-[125I]-iodophenyl)-tropane (β-CIT), sertraline potency was reduced by the I425V mutation but citalopram potency was unchanged. Mutation at the site of cGMP-dependent phosphorylation, Thr-276, decreased the potency of each of the drugs tested. This effect was also observed in studies with β-CIT where both citalopram and sertraline were less potent at displacing this high-affinity ligand. These results support an influence of Thr-276 on the conformation of inhibitor binding sites of serotonin transporter, and also suggest that the sertraline binding site contains unique determinants that are not shared with the other tested inhibitors.