Neurotransmitter sodium symporter
|Sodium:neurotransmitter symporter family|
Members of the Neurotransmitter:Sodium Symporter (NSS) family (TC# 2.A.22) catalyze uptake of a variety of neurotransmitters, amino acids, osmolytes and related nitrogenous substances by a solute:Na+ symport mechanism. The NSS family is a member of the APC Superfamily. Its constituents have been found in bacteria, archaea and eukaryotes.
Neurotransmitter transport systems are responsible for the release, re-uptake and recycling of neurotransmitters at synapses. High affinity transport proteins found in the plasma membrane of presynaptic nerve terminals and glial cells are responsible for the removal, from the extracellular space, of released-transmitters, thereby terminating their actions.
The majority of the transporters constitute an extensive family of homologous proteins that derive energy from the co-transport of Na+ and Cl−, in order to transport neurotransmitter molecules into the cell against their concentration gradient.
Since neurotransmitter:sodium symporters (NSS) have a critical role in regulating neurotransmission, they are targets for psychostimulants, anti-depressants and other drugs. In eukaryotic NSS, chloride is transported together with the neurotransmitter. However, transport by the bacterial homologues LeuT, Tyt1 and TnaT is chloride-independent. The crystal structure of LeuT ( In contrast to wild-type GAT-1, a marked stimulation of the rate of net flux (but not of exchange) was observed when the internal pH was lowered. Equivalent mutations introduced in the mouse GABA transporter GAT4 (SLC6A11) and the human dopamine transporter DAT (SLC6A3) also resulted in chloride-independent transport. The reciprocal mutations in LeuT and Tyt1 rendered substrate binding and/or uptake by these bacterial NSS chloride-dependent. Zomot et al.'s data indicated that the negative charge, provided either by chloride or by the transporter itself, is required during binding and translocation of the neurotransmitter, probably to counterbalance the charge of the co-transported sodium ions.) reveals an occluded binding pocket containing leucine and two sodium ions. Zomot et al., (2007) found that introduction of a negatively charged amino acid at or near one of the two putative sodium-binding sites of the GABA (γ-aminobutyric acid) transporter GAT-1 from rat brain (also called SLC6A1) renders both net flux and exchange of GABA largely chloride-independent.
The generalized transport reaction for the members of this family is:
- solute (out) + Na+ (out) → solute (in) + Na+ (in).
The family has a common structure of 12 presumed transmembrane helices and includes carriers for gamma-aminobutyric acid (GABA), noradrenaline/adrenaline, dopamine, serotonin, proline, glycine, choline, betaine, taurine and other small molecules.
NSS carriers are structurally distinct from the second more-restricted family of plasma membrane transporters, which are responsible for excitatory amino acid transport (see TC# 2.A.23). The latter couple glutamate and aspartate uptake to the cotransport of Na+ and the counter-transport of K+, with no apparent dependence on Cl−. In addition, both of these transporter families are distinct from the vesicular neurotransmitter transporters. Sequence analysis of the Na+/Cl− neurotransmitter superfamily reveals that it can be divided into four subfamilies, these being transporters for monoamines, the amino acids proline and glycine, GABA, and a group of orphan transporters.
Tavoulari et al. (2011) described conversion of the Cl− -independent prokaryotic tryptophan transporter TnaT (2.A.22.4.1) to a fully functional Cl− -dependent form by a single point mutation, D268S. Mutations in TnaT-D268S, in wild type TnaT and in a serotonin transporter (SERT; 2.A.22.1.1) provided direct evidence for the involvement of each of the proposed residues in Cl− coordination. In both SERT and TnaT-D268S, Cl− and Na+ mutually increase each other's potency, consistent with am electrostatic interaction through adjacent binding sites.
The crystal structure of a bacterial member of the NSS family has been determined complexed to leucine and 2 Na+. The protein core consists of the first ten of the 12 TMSs with segments 1-5 and 6-10 exhibiting a pseudo-2-fold axis of symmetry in the plane of the membrane. Leucine and the sodium ions are bound within the protein core, halfway across the membrane bilayer, in an occluded site devoid of water. The leucine and ion binding sites are defined by partially unwound transmembrane helices, with main-chain atoms and helix dipoles having key roles in substrate and ion binding. The binding pocket of LeuT contains two metal binding sites. The first ion in site NA1 is directly coupled to the bound substrate (Leu) with the second ion in the neighboring site (NA2) approximately 7 Å away. Double ion occupancy of the binding pocket is required to ensure substrate coupling to Na+ (but not to Li+ or K+ cations). The presence of the ion in site NA2 is required for structural stability of the binding pocket as well as amplified selectivity for Na+ in the case of double ion occupancy.
There are several crystal structures available for a couple members of the NSS family:
- 2.A.22.1.7 - Dopamine transporter: , , , ,
- 2.A.22.4.2 - The amino acid (leucine):2 Na+ symporter, LeuTAa: more) , , , , , , , (
Several characterized proteins are classified within the NSS family and can be found in the Transporter Classification Database.
- Betaine transporter (SLC6A12) InterPro: IPR002983
- Creatine transporter (SLC6A8) InterPro: IPR002984
- Dopamine neurotransmitter transporter (SLC6A3) InterPro: IPR002436
- Inebriated neurotransmitter transporter InterPro: IPR002944
- GABA neurotransmitter transporter GAT-1 (SLC6A1) InterPro: IPR002980
- GABA neurotransmitter transporter GAT-2 (SLC6A13) InterPro: IPR002981
- GABA neurotransmitter transporter GAT-3 (SLC6A11) InterPro: IPR002982
- Glycine neurotransmitter transporter, type 1 (SLC6A9) InterPro: IPR003028
- Noradrenaline neurotransmitter transporter (SLC6A9) InterPro: IPR002435
- Orphan neurotransmitter transporter (SLC6A15) InterPro: IPR002438
- Serotonin (5-HT) neurotransmitter transporter, N-terminal (SLC6A4) InterPro: IPR013086
- Taurine transporter (SLC6A6) InterPro: IPR002434
Human proteins containing this domain
- Transporter Classification Database (tcdb.org) for more detailed description of this family
- APC Superfamily
- Membrane transport proteins
- Rudnick, G; Krämer, R; Blakely, RD; Murphy, DL; Verrey, F (January 2014). "The SLC6 transporters: perspectives on structure, functions, regulation, and models for transporter dysfunction.". Pflugers Archive 466 (1): 25–42. doi:10.1007/s00424-013-1410-1. PMC 3930102. PMID 24337881.
- Saier, MH Jr. "2.A.22 The Neurotransmitter:Sodium Symporter (NSS) Family". Transporter Classification Database.
- Attwell D, Bouvier M (1992). "Cloners quick on the uptake". Curr. Biol. 2 (10): 541–543. doi:10.1016/0960-9822(92)90024-5. PMID 15336049.
- Zomot, E; Bendahan, A; Quick, M; Zhao, Y; Javitch, JA; Kanner, BI (October 11, 2007). "Mechanism of chloride interaction with neurotransmitter:sodium symporters.". Nature 449 (7163): 726–30. doi:10.1038/nature06133. PMID 17704762.
- Malandro MS, Kilberg MS (1996). "Molecular biology of mammalian amino acid transporters". Annu. Rev. Biochem. 65: 305–336. doi:10.1146/annurev.bi.65.070196.001513. PMID 8811182.
- Arriza JL, Amara SG (1993). "Neurotransmitter transporters: three distinct gene families". Curr. Opin. Neurobiol. 3 (3): 337–344. doi:10.1016/0959-4388(93)90126-J. PMID 8103691.
- Uhl GR, Johnson PS (1994). "Neurotransmitter transporters: three important gene families for neuronal function". J. Exp. Biol. 196: 229–236. PMID 7823024.
- Nelson N, Lill H (1998). "Homologies and family relationships among Na+/Cl− neurotransmitter transporters". Meth. Enzymol. Methods in Enzymology 296: 425–436. doi:10.1016/S0076-6879(98)96030-X. ISBN 978-0-12-182197-5. PMID 9779464.
- Tavoulari, S; Rizwan, AN; Forrest, LR; Rudnick, G (January 28, 2011). "Reconstructing a chloride-binding site in a bacterial neurotransmitter transporter homologue.". Journal of Biological Chemistry 286 (4): 2834–42. doi:10.1074/jbc.M110.186064. PMC 3024779. PMID 21115480.
- Yamashita, A; Singh, SK; Kawate, T; Jin, Y; Gouaux, E (September 8, 2005). "Crystal structure of a bacterial homologue of Na+/Cl--dependent neurotransmitter transporters.". Nature 437 (7056): 215–23. doi:10.1038/nature03978. PMID 16041361.
- Caplan, DA; Subbotina, JO; Noskov, SY (November 15, 2008). "Molecular mechanism of ion-ion and ion-substrate coupling in the Na+-dependent leucine transporter LeuT.". Biophysical Journal 95 (10): 4613–21. doi:10.1529/biophysj.108.139741. PMC 2576368. PMID 18708457.