ID   EAA4_MOUSE              Reviewed;         561 AA.
AC   O35544;
DT   15-JUL-1998, integrated into UniProtKB/Swiss-Prot.
DT   01-JAN-1998, sequence version 1.
DT   27-MAR-2024, entry version 163.
DE   RecName: Full=Excitatory amino acid transporter 4;
DE   AltName: Full=High-affinity neuronal glutamate transporter;
DE   AltName: Full=Sodium-dependent glutamate/aspartate transporter;
DE   AltName: Full=Solute carrier family 1 member 6;
GN   Name=Slc1a6; Synonyms=Eaat4;
OS   Mus musculus (Mouse).
OC   Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia;
OC   Eutheria; Euarchontoglires; Glires; Rodentia; Myomorpha; Muroidea; Muridae;
OC   Murinae; Mus; Mus.
OX   NCBI_TaxID=10090;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, SUBCELLULAR LOCATION,
RP   BIOPHYSICOCHEMICAL PROPERTIES, AND TISSUE SPECIFICITY.
RC   STRAIN=BALB/cJ; TISSUE=Cerebellum;
RX   PubMed=9379843; DOI=10.1016/s0169-328x(97)00169-1;
RA   Maeno-Hikichi Y., Tanaka K., Shibata T., Watanabe M., Inoue Y.,
RA   Mukainaka Y., Wada K.;
RT   "Structure and functional expression of the cloned mouse neuronal high-
RT   affinity glutamate transporter.";
RL   Brain Res. Mol. Brain Res. 48:176-180(1997).
RN   [2]
RP   IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
RC   TISSUE=Brain;
RX   PubMed=21183079; DOI=10.1016/j.cell.2010.12.001;
RA   Huttlin E.L., Jedrychowski M.P., Elias J.E., Goswami T., Rad R.,
RA   Beausoleil S.A., Villen J., Haas W., Sowa M.E., Gygi S.P.;
RT   "A tissue-specific atlas of mouse protein phosphorylation and expression.";
RL   Cell 143:1174-1189(2010).
CC   -!- FUNCTION: Sodium-dependent, high-affinity amino acid transporter that
CC       mediates the uptake of L-glutamate and also L-aspartate and D-aspartate
CC       (PubMed:9379843). Functions as a symporter that transports one amino
CC       acid molecule together with two or three Na(+) ions and one proton, in
CC       parallel with the counter-transport of one K(+) ion. Mediates Cl(-)
CC       flux that is not coupled to amino acid transport; this avoids the
CC       accumulation of negative charges due to aspartate and Na(+) symport (By
CC       similarity). Plays a redundant role in the rapid removal of released
CC       glutamate from the synaptic cleft, which is essential for terminating
CC       the postsynaptic action of glutamate (Probable).
CC       {ECO:0000250|UniProtKB:O35921, ECO:0000269|PubMed:9379843,
CC       ECO:0000305}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=H(+)(out) + K(+)(in) + L-glutamate(out) + 3 Na(+)(out) =
CC         H(+)(in) + K(+)(out) + L-glutamate(in) + 3 Na(+)(in);
CC         Xref=Rhea:RHEA:70699, ChEBI:CHEBI:15378, ChEBI:CHEBI:29101,
CC         ChEBI:CHEBI:29103, ChEBI:CHEBI:29985;
CC         Evidence={ECO:0000250|UniProtKB:P48664};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=H(+)(out) + K(+)(in) + L-aspartate(out) + 3 Na(+)(out) =
CC         H(+)(in) + K(+)(out) + L-aspartate(in) + 3 Na(+)(in);
CC         Xref=Rhea:RHEA:70851, ChEBI:CHEBI:15378, ChEBI:CHEBI:29101,
CC         ChEBI:CHEBI:29103, ChEBI:CHEBI:29991;
CC         Evidence={ECO:0000250|UniProtKB:P48664};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=D-aspartate(out) + H(+)(out) + K(+)(in) + 3 Na(+)(out) = D-
CC         aspartate(in) + H(+)(in) + K(+)(out) + 3 Na(+)(in);
CC         Xref=Rhea:RHEA:71379, ChEBI:CHEBI:15378, ChEBI:CHEBI:29101,
CC         ChEBI:CHEBI:29103, ChEBI:CHEBI:29990;
CC         Evidence={ECO:0000250|UniProtKB:P48664};
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=40 uM for L-glutamate {ECO:0000269|PubMed:9379843};
CC   -!- SUBUNIT: Homotrimer. {ECO:0000305}.
CC   -!- SUBCELLULAR LOCATION: Cell membrane {ECO:0000269|PubMed:9379843};
CC       Multi-pass membrane protein {ECO:0000305}.
CC   -!- TISSUE SPECIFICITY: Brain specific. {ECO:0000269|PubMed:9379843}.
CC   -!- DOMAIN: Contains eight transmembrane regions plus two helical hairpins
CC       that dip into the membrane. These helical hairpin structures play an
CC       important role in the transport process. The first enters the membrane
CC       from the cytoplasmic side, the second one from the extracellular side.
CC       During the transport cycle, the regions involved in amino acid
CC       transport, and especially the helical hairpins, move vertically by
CC       about 15-18 Angstroms, alternating between exposure to the aqueous
CC       phase and reinsertion in the lipid bilayer. In contrast, the regions
CC       involved in trimerization do not move. {ECO:0000250|UniProtKB:P43003}.
CC   -!- SIMILARITY: Belongs to the dicarboxylate/amino acid:cation symporter
CC       (DAACS) (TC 2.A.23) family. SLC1A6 subfamily. {ECO:0000305}.
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DR   EMBL; D83262; BAA23640.1; -; mRNA.
DR   CCDS; CCDS23970.1; -.
DR   RefSeq; NP_033226.1; NM_009200.3.
DR   AlphaFoldDB; O35544; -.
DR   SMR; O35544; -.
DR   BioGRID; 203292; 1.
DR   STRING; 10090.ENSMUSP00000005490; -.
DR   GlyCosmos; O35544; 3 sites, No reported glycans.
DR   GlyGen; O35544; 3 sites.
DR   iPTMnet; O35544; -.
DR   PhosphoSitePlus; O35544; -.
DR   SwissPalm; O35544; -.
DR   MaxQB; O35544; -.
DR   PaxDb; 10090-ENSMUSP00000005490; -.
DR   ProteomicsDB; 275426; -.
DR   Antibodypedia; 26923; 203 antibodies from 25 providers.
DR   DNASU; 20513; -.
DR   Ensembl; ENSMUST00000005490.10; ENSMUSP00000005490.9; ENSMUSG00000005357.10.
DR   GeneID; 20513; -.
DR   KEGG; mmu:20513; -.
DR   UCSC; uc007fyg.2; mouse.
DR   AGR; MGI:1096331; -.
DR   CTD; 6511; -.
DR   MGI; MGI:1096331; Slc1a6.
DR   VEuPathDB; HostDB:ENSMUSG00000005357; -.
DR   eggNOG; KOG3787; Eukaryota.
DR   GeneTree; ENSGT00940000159972; -.
DR   HOGENOM; CLU_019375_3_2_1; -.
DR   InParanoid; O35544; -.
DR   OMA; VGDIFIH; -.
DR   OrthoDB; 49426at2759; -.
DR   PhylomeDB; O35544; -.
DR   TreeFam; TF315206; -.
DR   Reactome; R-MMU-210500; Glutamate Neurotransmitter Release Cycle.
DR   Reactome; R-MMU-425393; Transport of inorganic cations/anions and amino acids/oligopeptides.
DR   BioGRID-ORCS; 20513; 2 hits in 75 CRISPR screens.
DR   ChiTaRS; Slc1a6; mouse.
DR   PRO; PR:O35544; -.
DR   Proteomes; UP000000589; Chromosome 10.
DR   RNAct; O35544; Protein.
DR   Bgee; ENSMUSG00000005357; Expressed in cerebellar vermis and 70 other cell types or tissues.
DR   ExpressionAtlas; O35544; baseline and differential.
DR   GO; GO:0098978; C:glutamatergic synapse; IDA:SynGO.
DR   GO; GO:0005794; C:Golgi apparatus; IDA:MGI.
DR   GO; GO:0045111; C:intermediate filament cytoskeleton; ISO:MGI.
DR   GO; GO:0016020; C:membrane; IDA:MGI.
DR   GO; GO:0098796; C:membrane protein complex; ISO:MGI.
DR   GO; GO:0098688; C:parallel fiber to Purkinje cell synapse; ISO:MGI.
DR   GO; GO:0005886; C:plasma membrane; ISS:UniProtKB.
DR   GO; GO:0045211; C:postsynaptic membrane; ISO:MGI.
DR   GO; GO:0042734; C:presynaptic membrane; IDA:SynGO.
DR   GO; GO:0015501; F:glutamate:sodium symporter activity; ISS:UniProtKB.
DR   GO; GO:0005314; F:high-affinity L-glutamate transmembrane transporter activity; ISS:UniProtKB.
DR   GO; GO:0015183; F:L-aspartate transmembrane transporter activity; ISS:UniProtKB.
DR   GO; GO:0005313; F:L-glutamate transmembrane transporter activity; IMP:MGI.
DR   GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR   GO; GO:0008509; F:monoatomic anion transmembrane transporter activity; ISO:MGI.
DR   GO; GO:0015175; F:neutral L-amino acid transmembrane transporter activity; IBA:GO_Central.
DR   GO; GO:0051649; P:establishment of localization in cell; IMP:MGI.
DR   GO; GO:0140009; P:L-aspartate import across plasma membrane; ISS:UniProtKB.
DR   GO; GO:0098712; P:L-glutamate import across plasma membrane; ISS:UniProtKB.
DR   GO; GO:0015813; P:L-glutamate transmembrane transport; IMP:MGI.
DR   GO; GO:0001504; P:neurotransmitter uptake; ISO:MGI.
DR   GO; GO:0042391; P:regulation of membrane potential; IMP:MGI.
DR   Gene3D; 1.10.3860.10; Sodium:dicarboxylate symporter; 1.
DR   InterPro; IPR001991; Na-dicarboxylate_symporter.
DR   InterPro; IPR018107; Na-dicarboxylate_symporter_CS.
DR   InterPro; IPR036458; Na:dicarbo_symporter_sf.
DR   PANTHER; PTHR11958:SF67; EXCITATORY AMINO ACID TRANSPORTER 4; 1.
DR   PANTHER; PTHR11958; SODIUM/DICARBOXYLATE SYMPORTER-RELATED; 1.
DR   Pfam; PF00375; SDF; 1.
DR   PRINTS; PR00173; EDTRNSPORT.
DR   SUPFAM; SSF118215; Proton glutamate symport protein; 1.
DR   PROSITE; PS00713; NA_DICARBOXYL_SYMP_1; 1.
DR   PROSITE; PS00714; NA_DICARBOXYL_SYMP_2; 1.
DR   Genevisible; O35544; MM.
PE   1: Evidence at protein level;
KW   Amino-acid transport; Cell membrane; Chloride; Glycoprotein; Membrane;
KW   Metal-binding; Phosphoprotein; Potassium; Reference proteome; Sodium;
KW   Symport; Transmembrane; Transmembrane helix; Transport.
FT   CHAIN           1..561
FT                   /note="Excitatory amino acid transporter 4"
FT                   /id="PRO_0000202071"
FT   TOPO_DOM        1..52
FT                   /note="Cytoplasmic"
FT                   /evidence="ECO:0000255"
FT   TRANSMEM        53..73
FT                   /note="Helical"
FT                   /evidence="ECO:0000255"
FT   TRANSMEM        96..116
FT                   /note="Helical"
FT                   /evidence="ECO:0000255"
FT   TRANSMEM        130..150
FT                   /note="Helical"
FT                   /evidence="ECO:0000255"
FT   TRANSMEM        259..282
FT                   /note="Helical; Name=4"
FT                   /evidence="ECO:0000250|UniProtKB:P43003"
FT   TRANSMEM        292..319
FT                   /note="Helical; Name=5"
FT                   /evidence="ECO:0000250|UniProtKB:P43003"
FT   TRANSMEM        341..362
FT                   /note="Helical; Name=6"
FT                   /evidence="ECO:0000250|UniProtKB:P43003"
FT   INTRAMEM        368..398
FT                   /note="Discontinuously helical"
FT                   /evidence="ECO:0000250|UniProtKB:P43003"
FT   TRANSMEM        408..434
FT                   /note="Helical; Name=7"
FT                   /evidence="ECO:0000250|UniProtKB:P43003"
FT   INTRAMEM        448..481
FT                   /note="Discontinuously helical"
FT                   /evidence="ECO:0000250|UniProtKB:P43003"
FT   TRANSMEM        495..516
FT                   /note="Helical; Name=8"
FT                   /evidence="ECO:0000250|UniProtKB:P43003"
FT   BINDING         385..387
FT                   /ligand="L-aspartate"
FT                   /ligand_id="ChEBI:CHEBI:29991"
FT                   /evidence="ECO:0000250|UniProtKB:P43003"
FT   BINDING         416
FT                   /ligand="Na(+)"
FT                   /ligand_id="ChEBI:CHEBI:29101"
FT                   /ligand_label="1"
FT                   /evidence="ECO:0000250|UniProtKB:O59010"
FT   BINDING         418
FT                   /ligand="Na(+)"
FT                   /ligand_id="ChEBI:CHEBI:29101"
FT                   /ligand_label="2"
FT                   /evidence="ECO:0000250|UniProtKB:P43003"
FT   BINDING         420
FT                   /ligand="Na(+)"
FT                   /ligand_id="ChEBI:CHEBI:29101"
FT                   /ligand_label="1"
FT                   /evidence="ECO:0000250|UniProtKB:O59010"
FT   BINDING         424
FT                   /ligand="L-aspartate"
FT                   /ligand_id="ChEBI:CHEBI:29991"
FT                   /evidence="ECO:0000250|UniProtKB:P43003"
FT   BINDING         465..469
FT                   /ligand="L-aspartate"
FT                   /ligand_id="ChEBI:CHEBI:29991"
FT                   /evidence="ECO:0000250|UniProtKB:P43003"
FT   BINDING         498
FT                   /ligand="L-aspartate"
FT                   /ligand_id="ChEBI:CHEBI:29991"
FT                   /evidence="ECO:0000250|UniProtKB:P43003"
FT   BINDING         505
FT                   /ligand="L-aspartate"
FT                   /ligand_id="ChEBI:CHEBI:29991"
FT                   /evidence="ECO:0000250|UniProtKB:P43003"
FT   BINDING         505
FT                   /ligand="Na(+)"
FT                   /ligand_id="ChEBI:CHEBI:29101"
FT                   /ligand_label="1"
FT                   /evidence="ECO:0000250|UniProtKB:O59010"
FT   BINDING         509
FT                   /ligand="Na(+)"
FT                   /ligand_id="ChEBI:CHEBI:29101"
FT                   /ligand_label="1"
FT                   /evidence="ECO:0000250|UniProtKB:O59010"
FT   MOD_RES         2
FT                   /note="Phosphoserine"
FT                   /evidence="ECO:0000250|UniProtKB:O35921"
FT   CARBOHYD        213
FT                   /note="N-linked (GlcNAc...) asparagine"
FT                   /evidence="ECO:0000255"
FT   CARBOHYD        229
FT                   /note="N-linked (GlcNAc...) asparagine"
FT                   /evidence="ECO:0000255"
FT   CARBOHYD        236
FT                   /note="N-linked (GlcNAc...) asparagine"
FT                   /evidence="ECO:0000255"
SQ   SEQUENCE   561 AA;  60784 MW;  CCFE3FD499D4CBBB CRC64;
     MSSHGNSLFL RESGAGGGCL QGLQDSLQQR ALRTRLRLQT MTREHVRRFL RRNAFILLTV
     SAVIIGVSLA FALRPYQLSY RQIKYFSFPG ELLMRMLQML VLPLIVSSLV TGMASLDNKA
     TGRMGMRAAV YYMVTTVIAV FIGILMVTII HPGKGSKEGL HREGRIETVP TADAFMDLVR
     NMFPPNLVEA CFKQFKTQYS TRVVTRTIVR TDNGSELGAS MSPTSSVENE TSILENVTRA
     LGTLQEVISF EETVPVPGSA NGINALGLVV FSVAFGLVIG GMKHKGRVLR DFFDSLNEAI
     MRLVGIIIWY APVGILFLIA GKILEMEDMA VLGGQLGMYT LTVIVGLFLH AGGVLPLIYF
     LVTHRNPFPF IGGMLQALIT AMGTSSSSAT LPITFRCLEE GLGVDRRITR FVLPVGATVN
     MDGTALYEAL AAIFIAQVNN YELNLGQITT ISITATAASV GAAGIPQAGL VTMVIVLTSV
     GLPTEDITLI IAVDWFLDRL RTMTNVLGDS IGAAVIEHLS QRELELQEAE LTLPSLGKPY
     KSLMAQEKGA SRGRGGNESV M
//