The cytoplasmic substrate permeation pathway of serotonin transporter.
Serotonin transporter (SERT) catalyzes reuptake of the neurotransmitter serotonin (5-HT) and is a target for antidepressant drugs and psychostimulants. It is a member of a large family of neurotransmitter and amino acid transporters. A recent study using site-directed cysteine modification identified a helical region of the transporter with high accessibility to the cytoplasm. Subsequently, the high resolution structure of LeuT, a prokaryotic homologue, showed that the residues corresponding to this helical region are part of the fifth transmembrane domain. The accessibility of these positions is now shown to depend on conformational changes corresponding to interconversion of SERT between two forms that face the extracellular medium and the cytoplasm, respectively. Binding of the extracellular inhibitor cocaine decreased accessibility at these positions, whereas 5-HT, the transported substrate, increased it. The effect of 5-HT required the simultaneous presence of Na+ and Cl-, which are transported into the cell together (symported) with 5-HT. In light of the LeuT structure, these results begin to define the pathway through which 5-HT diffuses between its binding site and the cytoplasm. They also confirm a prediction of the alternating access model for transport, namely, that all symported substrates must bind together before translocation.
Zhang
Yuan-Wei
YW
Department of Pharmacology, Yale University School of Medicine, New Haven, Connecticut 06520-8066, USA.
Rudnick
Gary
G
D000818:Animals; D002712:Chlorides / Q000737:chemistry; D003042:Cocaine / Q000494:pharmacology; D003593:Cytoplasm / Q000378:metabolism; D018765:Dopamine Uptake Inhibitors / Q000494:pharmacology; D005020:Ethyl Methanesulfonate / Q000031:analogs & derivatives / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D011487:Protein Conformation; D021381:Protein Transport; D051381:Rats; D012701:Serotonin / Q000737:chemistry*; D050486:Serotonin Plasma Membrane Transport Proteins / Q000737:chemistry / Q000502:physiology*; D012964:Sodium / Q000737:chemistry; D012965:Sodium Chloride / Q000494:pharmacology
D002712:Chlorides; D018765:Dopamine Uptake Inhibitors; D050486:Serotonin Plasma Membrane Transport Proteins; C093171:methanethiosulfonate ethylammonium; D012701:Serotonin; D012965:Sodium Chloride; D005020:Ethyl Methanesulfonate; D012964:Sodium; D003042:Cocaine