Elsevier

Methods in Enzymology

Volume 296, 1998, Pages 331-346
Methods in Enzymology

[23] Probing structure of neurotransmitter transporters by substituted-cysteine accessibility method

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Publisher Summary

Neurotransmitter reuptake by transport proteins is a major mechanism for terminating synaptic transmission. These transporters operate by coupling the net electrochemical gradient for sodium, chloride, and neurotransmitter to the transmembrane translocation of these substrates. Such a transporter must contain a water-accessible transport pathway lined by residues from the membrane-spanning segments. Neurotransmitter, sodium, and chloride must bind to specific sites within this pathway and cause a conformational change that alters the exposure of the bound substrates from the extracellular milieu to the intracellular milieu and reduces their binding affinity to facilitate intracellular release. Although the transport pathway bears some functional resemblance to an ion channel, the transporter gating mechanism must be considerably more complex. Likewise, the presence of binding sites for neurotransmitter and ions in membrane-spanning segments is reminiscent of the G-protein-coupled receptors but substrate translocation requires a mechanism by which structural changes in the transporter alter the exposure of the binding sites and their affinities. Substituted-cysteine accessibility method is described in the chapter.

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