ID   VIAAT_RAT               Reviewed;         525 AA.
AC   O35458;
DT   28-MAR-2003, integrated into UniProtKB/Swiss-Prot.
DT   01-JAN-1998, sequence version 1.
DT   27-MAR-2024, entry version 151.
DE   RecName: Full=Vesicular inhibitory amino acid transporter;
DE   AltName: Full=GABA and glycine transporter;
DE   AltName: Full=Solute carrier family 32 member 1;
DE   AltName: Full=Vesicular GABA transporter {ECO:0000303|PubMed:9349821};
DE            Short=rGVAT;
DE   AltName: Full=rat UNC-47 homolog {ECO:0000303|PubMed:9349821};
DE            Short=RUNC-47 {ECO:0000303|PubMed:9349821};
GN   Name=Slc32a1 {ECO:0000312|RGD:621402}; Synonyms=Vgat, Viaat;
OS   Rattus norvegicus (Rat).
OC   Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia;
OC   Eutheria; Euarchontoglires; Glires; Rodentia; Myomorpha; Muroidea; Muridae;
OC   Murinae; Rattus.
OX   NCBI_TaxID=10116;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [MRNA], TISSUE SPECIFICITY, FUNCTION, TRANSPORTER
RP   ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RC   STRAIN=Sprague-Dawley;
RX   PubMed=9349821; DOI=10.1038/39908;
RA   McIntire S.L., Reimer R.J., Schuske K., Edwards R.H., Jorgensen E.M.;
RT   "Identification and characterization of the vesicular GABA transporter.";
RL   Nature 389:870-876(1997).
RN   [2]
RP   SUBCELLULAR LOCATION, AND TISSUE SPECIFICITY.
RX   PubMed=9822734; DOI=10.1523/jneurosci.18-23-09733.1998;
RA   Chaudhry F.A., Reimer R.J., Bellocchio E.E., Danbolt N.C., Osen K.K.,
RA   Edwards R.H., Storm-Mathisen J.;
RT   "The vesicular GABA transporter, VGAT, localizes to synaptic vesicles in
RT   sets of glycinergic as well as GABAergic neurons.";
RL   J. Neurosci. 18:9733-9750(1998).
RN   [3]
RP   SUBCELLULAR LOCATION.
RX   PubMed=10036231; DOI=10.1242/jcs.112.6.811;
RA   Dumoulin A., Rostaing P., Bedet C., Levi S., Isambert M.F., Henry J.P.,
RA   Triller A., Gasnier B.;
RT   "Presence of the vesicular inhibitory amino acid transporter in GABAergic
RT   and glycinergic synaptic terminal boutons.";
RL   J. Cell Sci. 112:811-823(1999).
RN   [4]
RP   CHARACTERIZATION, AND TISSUE SPECIFICITY.
RC   TISSUE=Brain;
RX   PubMed=12031963; DOI=10.2337/diabetes.51.6.1763;
RA   Chessler S.D., Simonson W.T., Sweet I.R., Hammerle L.P.;
RT   "Expression of the vesicular inhibitory amino acid transporter in
RT   pancreatic islet cells: distribution of the transporter within rat
RT   islets.";
RL   Diabetes 51:1763-1771(2002).
RN   [5]
RP   TOPOLOGY.
RX   PubMed=19052203; DOI=10.1523/jneurosci.3887-08.2008;
RA   Martens H., Weston M.C., Boulland J.L., Groenborg M., Grosche J., Kacza J.,
RA   Hoffmann A., Matteoli M., Takamori S., Harkany T., Chaudhry F.A.,
RA   Rosenmund C., Erck C., Jahn R., Haertig W.;
RT   "Unique luminal localization of VGAT-C terminus allows for selective
RT   labeling of active cortical GABAergic synapses.";
RL   J. Neurosci. 28:13125-13131(2008).
RN   [6]
RP   FUNCTION, BIOPHYSICOCHEMICAL PROPERTIES, CAUTION, AND MUTAGENESIS OF
RP   GLU-213.
RX   PubMed=19843525; DOI=10.1074/jbc.m109.062414;
RA   Juge N., Muroyama A., Hiasa M., Omote H., Moriyama Y.;
RT   "Vesicular inhibitory amino acid transporter is a Cl-/gamma-aminobutyrate
RT   Co-transporter.";
RL   J. Biol. Chem. 284:35073-35078(2009).
RN   [7]
RP   FUNCTION, CAUTION, MUTAGENESIS OF GLU-213 AND LYS-351, BIOPHYSICOCHEMICAL
RP   PROPERTIES, AND TRANSPORTER ACTIVITY.
RX   PubMed=23919636; DOI=10.1111/jnc.12393;
RA   Juge N., Omote H., Moriyama Y.;
RT   "Vesicular GABA transporter (VGAT) transports beta-alanine.";
RL   J. Neurochem. 127:482-486(2013).
RN   [8]
RP   IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
RX   PubMed=22673903; DOI=10.1038/ncomms1871;
RA   Lundby A., Secher A., Lage K., Nordsborg N.B., Dmytriyev A., Lundby C.,
RA   Olsen J.V.;
RT   "Quantitative maps of protein phosphorylation sites across 14 different rat
RT   organs and tissues.";
RL   Nat. Commun. 3:876-876(2012).
CC   -!- FUNCTION: Antiporter that exchanges vesicular protons for cytosolic 4-
CC       aminobutanoate or to a lesser extend glycine, thus allowing their
CC       secretion from nerve terminals (By similarity) (PubMed:9349821). The
CC       transport is equally dependent on the chemical and electrical
CC       components of the proton gradient (PubMed:9349821). May also transport
CC       beta-alanine (PubMed:23919636). Acidification of GABAergic synaptic
CC       vesicles is a prerequisite for 4-aminobutanoate uptake (By similarity).
CC       {ECO:0000250|UniProtKB:O35633, ECO:0000269|PubMed:23919636,
CC       ECO:0000269|PubMed:9349821}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=beta-alanine(out) + n H(+)(in) = beta-alanine(in) + n
CC         H(+)(out); Xref=Rhea:RHEA:70987, ChEBI:CHEBI:15378,
CC         ChEBI:CHEBI:57966; Evidence={ECO:0000305|PubMed:23919636};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=4-aminobutanoate(out) + n H(+)(in) = 4-aminobutanoate(in) + n
CC         H(+)(out); Xref=Rhea:RHEA:70979, ChEBI:CHEBI:15378,
CC         ChEBI:CHEBI:59888; Evidence={ECO:0000269|PubMed:9349821};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=glycine(out) + n H(+)(in) = glycine(in) + n H(+)(out);
CC         Xref=Rhea:RHEA:70983, ChEBI:CHEBI:15378, ChEBI:CHEBI:57305;
CC         Evidence={ECO:0000250|UniProtKB:O35633};
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=5 mM for 4-aminobutanoate {ECO:0000269|PubMed:9349821};
CC         KM=3.5 mM for beta-alanine {ECO:0000269|PubMed:23919636};
CC         KM=0.8 mM for 4-aminobutanoate {ECO:0000269|PubMed:19843525};
CC         KM=2.1 mM for chloride {ECO:0000269|PubMed:19843525};
CC         KM=2.3 mM for chloride (for 4-aminobutanoate uptake)
CC         {ECO:0000269|PubMed:19843525};
CC         Vmax=38 nmol/min/mg enzyme toward beta-alanine
CC         {ECO:0000269|PubMed:23919636};
CC         Vmax=41 nmol/min/mg enzyme toward 4-aminobutanoate
CC         {ECO:0000269|PubMed:19843525};
CC         Vmax=90.6 nmol/min/mg enzyme toward chloride
CC         {ECO:0000269|PubMed:19843525};
CC         Vmax=7.4 nmol/min/mg enzyme toward chloride (for 4-aminobutanoate
CC         uptake) {ECO:0000269|PubMed:19843525};
CC   -!- SUBCELLULAR LOCATION: Cytoplasmic vesicle, secretory vesicle, synaptic
CC       vesicle membrane {ECO:0000269|PubMed:10036231,
CC       ECO:0000269|PubMed:9822734}; Multi-pass membrane protein {ECO:0000255}.
CC       Presynapse {ECO:0000269|PubMed:10036231}. Note=Presents in glycine-,
CC       GABA- or GABA- and glycine-containing boutons.
CC       {ECO:0000269|PubMed:10036231}.
CC   -!- TISSUE SPECIFICITY: Brain (PubMed:9822734, PubMed:9349821). Expressed
CC       at high levels within the neocortex, hippocampus, cerebellum, striatum,
CC       septal nuclei and the reticular nucleus of the thalamus
CC       (PubMed:9349821). Also expressed in islets where it is more abundant in
CC       the peripheral/mantle region (PubMed:12031963). Highly expressed in the
CC       nerve endings of GABA neurons in the brain and spinal cord but also in
CC       glycinergic nerve endings (PubMed:9822734). Expressed in glycine-,
CC       GABA- or GABA- and glycine-containing boutons (PubMed:10036231,
CC       PubMed:9822734). {ECO:0000269|PubMed:10036231,
CC       ECO:0000269|PubMed:12031963, ECO:0000269|PubMed:9349821,
CC       ECO:0000269|PubMed:9822734}.
CC   -!- SIMILARITY: Belongs to the amino acid/polyamine transporter 2 family.
CC       {ECO:0000305}.
CC   -!- CAUTION: Juge et al. shows that SLC32A1 is a symporter of both 4-
CC       aminobutanoate or glycine or beta-alanine with Cl(-) that operates
CC       according an electrical gradient without the need for a chemical
CC       gradient (PubMed:19843525, PubMed:23919636). However Farsi et al. and
CC       Egashira et al. confirm that SLC32A1 is an antiporter that exchanges
CC       vesicular protons for cytosolic 4-aminobutanoate or glycine and exclude
CC       any coupling with chloride (By similarity).
CC       {ECO:0000250|UniProtKB:O35633, ECO:0000269|PubMed:19843525,
CC       ECO:0000269|PubMed:23919636}.
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DR   EMBL; AF030253; AAB82950.1; -; mRNA.
DR   RefSeq; NP_113970.1; NM_031782.1.
DR   RefSeq; XP_006235503.1; XM_006235441.3.
DR   AlphaFoldDB; O35458; -.
DR   SMR; O35458; -.
DR   BioGRID; 249777; 2.
DR   IntAct; O35458; 2.
DR   MINT; O35458; -.
DR   STRING; 10116.ENSRNOP00000020720; -.
DR   PhosphoSitePlus; O35458; -.
DR   SwissPalm; O35458; -.
DR   PaxDb; 10116-ENSRNOP00000020720; -.
DR   ABCD; O35458; 2 sequenced antibodies.
DR   Ensembl; ENSRNOT00000020720.7; ENSRNOP00000020720.5; ENSRNOG00000015393.7.
DR   Ensembl; ENSRNOT00055047459; ENSRNOP00055039036; ENSRNOG00055027414.
DR   Ensembl; ENSRNOT00060046377; ENSRNOP00060038540; ENSRNOG00060026774.
DR   Ensembl; ENSRNOT00065048724; ENSRNOP00065039956; ENSRNOG00065028288.
DR   GeneID; 83612; -.
DR   KEGG; rno:83612; -.
DR   UCSC; RGD:621402; rat.
DR   AGR; RGD:621402; -.
DR   CTD; 140679; -.
DR   RGD; 621402; Slc32a1.
DR   eggNOG; KOG4303; Eukaryota.
DR   GeneTree; ENSGT00490000043380; -.
DR   HOGENOM; CLU_036432_0_0_1; -.
DR   InParanoid; O35458; -.
DR   OMA; RSWMMLI; -.
DR   OrthoDB; 2908005at2759; -.
DR   PhylomeDB; O35458; -.
DR   TreeFam; TF312818; -.
DR   Reactome; R-RNO-425393; Transport of inorganic cations/anions and amino acids/oligopeptides.
DR   Reactome; R-RNO-888590; GABA synthesis, release, reuptake and degradation.
DR   PRO; PR:O35458; -.
DR   Proteomes; UP000002494; Chromosome 3.
DR   Bgee; ENSRNOG00000015393; Expressed in cerebellum and 2 other cell types or tissues.
DR   GO; GO:0009986; C:cell surface; ISO:RGD.
DR   GO; GO:0051286; C:cell tip; ISO:RGD.
DR   GO; GO:0044316; C:cone cell pedicle; ISO:RGD.
DR   GO; GO:0030425; C:dendrite; ISO:RGD.
DR   GO; GO:0044292; C:dendrite terminus; ISO:RGD.
DR   GO; GO:0098982; C:GABA-ergic synapse; IDA:SynGO.
DR   GO; GO:0060077; C:inhibitory synapse; IDA:BHF-UCL.
DR   GO; GO:0043005; C:neuron projection; ISO:RGD.
DR   GO; GO:0044306; C:neuron projection terminus; ISO:RGD.
DR   GO; GO:0098793; C:presynapse; IDA:UniProtKB.
DR   GO; GO:0048786; C:presynaptic active zone; ISO:RGD.
DR   GO; GO:0045202; C:synapse; IDA:MGI.
DR   GO; GO:0008021; C:synaptic vesicle; IDA:UniProtKB.
DR   GO; GO:0030672; C:synaptic vesicle membrane; IDA:SynGO.
DR   GO; GO:0015185; F:gamma-aminobutyric acid transmembrane transporter activity; IMP:UniProtKB.
DR   GO; GO:0140800; F:gamma-aminobutyric acid:proton antiporter activity; IDA:UniProtKB.
DR   GO; GO:0015187; F:glycine transmembrane transporter activity; IMP:UniProtKB.
DR   GO; GO:0140799; F:glycine:proton antiporter activity; ISS:UniProtKB.
DR   GO; GO:0001762; P:beta-alanine transport; IDA:UniProtKB.
DR   GO; GO:0051939; P:gamma-aminobutyric acid import; IMP:UniProtKB.
DR   GO; GO:0015812; P:gamma-aminobutyric acid transport; IDA:BHF-UCL.
DR   GO; GO:0015816; P:glycine transport; IMP:UniProtKB.
DR   GO; GO:0021766; P:hippocampus development; IEP:RGD.
DR   GO; GO:0098700; P:neurotransmitter loading into synaptic vesicle; IDA:SynGO.
DR   GO; GO:0006836; P:neurotransmitter transport; TAS:RGD.
DR   InterPro; IPR013057; AA_transpt_TM.
DR   PANTHER; PTHR22950; AMINO ACID TRANSPORTER; 1.
DR   PANTHER; PTHR22950:SF689; VESICULAR INHIBITORY AMINO ACID TRANSPORTER; 1.
DR   Pfam; PF01490; Aa_trans; 1.
DR   Genevisible; O35458; RN.
PE   1: Evidence at protein level;
KW   Cell projection; Cytoplasmic vesicle; Membrane; Neurotransmitter transport;
KW   Nitration; Reference proteome; Synapse; Transmembrane; Transmembrane helix;
KW   Transport.
FT   CHAIN           1..525
FT                   /note="Vesicular inhibitory amino acid transporter"
FT                   /id="PRO_0000093823"
FT   TOPO_DOM        1..132
FT                   /note="Cytoplasmic"
FT                   /evidence="ECO:0000305|PubMed:19052203"
FT   TRANSMEM        133..153
FT                   /note="Helical"
FT                   /evidence="ECO:0000255"
FT   TOPO_DOM        154..204
FT                   /note="Lumenal, vesicle"
FT                   /evidence="ECO:0000305|PubMed:19052203"
FT   TRANSMEM        205..225
FT                   /note="Helical"
FT                   /evidence="ECO:0000255"
FT   TOPO_DOM        226..265
FT                   /note="Cytoplasmic"
FT                   /evidence="ECO:0000305|PubMed:19052203"
FT   TRANSMEM        266..286
FT                   /note="Helical"
FT                   /evidence="ECO:0000255"
FT   TOPO_DOM        287..305
FT                   /note="Lumenal, vesicle"
FT                   /evidence="ECO:0000305|PubMed:19052203"
FT   TRANSMEM        306..326
FT                   /note="Helical"
FT                   /evidence="ECO:0000255"
FT   TOPO_DOM        327..341
FT                   /note="Cytoplasmic"
FT                   /evidence="ECO:0000305|PubMed:19052203"
FT   TRANSMEM        342..362
FT                   /note="Helical"
FT                   /evidence="ECO:0000255"
FT   TOPO_DOM        363..383
FT                   /note="Lumenal, vesicle"
FT                   /evidence="ECO:0000305|PubMed:19052203"
FT   TRANSMEM        384..404
FT                   /note="Helical"
FT                   /evidence="ECO:0000255"
FT   TOPO_DOM        405..438
FT                   /note="Cytoplasmic"
FT                   /evidence="ECO:0000305|PubMed:19052203"
FT   TRANSMEM        439..459
FT                   /note="Helical"
FT                   /evidence="ECO:0000255"
FT   TOPO_DOM        460..461
FT                   /note="Lumenal, vesicle"
FT                   /evidence="ECO:0000305|PubMed:19052203"
FT   TRANSMEM        462..482
FT                   /note="Helical"
FT                   /evidence="ECO:0000255"
FT   TOPO_DOM        483..489
FT                   /note="Cytoplasmic"
FT                   /evidence="ECO:0000305|PubMed:19052203"
FT   TRANSMEM        490..510
FT                   /note="Helical"
FT                   /evidence="ECO:0000255"
FT   TOPO_DOM        511..525
FT                   /note="Lumenal, vesicle"
FT                   /evidence="ECO:0000305|PubMed:19052203"
FT   MOD_RES         186
FT                   /note="3'-nitrotyrosine"
FT                   /evidence="ECO:0000250|UniProtKB:O35633"
FT   MUTAGEN         213
FT                   /note="E->A: Loss of GABA and glycine transport activity.
FT                   Loss of potential gradient-induced Cl(-) uptake. Loss of
FT                   beta-alanine transport activity."
FT                   /evidence="ECO:0000269|PubMed:19843525,
FT                   ECO:0000269|PubMed:23919636"
FT   MUTAGEN         351
FT                   /note="K->A: Retains around 80% of the beta-alanine
FT                   transport activity."
FT                   /evidence="ECO:0000269|PubMed:23919636"
SQ   SEQUENCE   525 AA;  57407 MW;  33C5E8D31B7BD510 CRC64;
     MATLLRSKLT NVATSVSNKS QAKVSGMFAR MGFQAATDEE AVGFAHCDDL DFEHRQGLQM
     DILKSEGEPC GDEGAEPPVE GDIHYQRGGA PLPPSGSKDQ AVGAGGEFGG HDKPKITAWE
     AGWNVTNAIQ GMFVLGLPYA ILHGGYLGLF LIIFAAVVCC YTGKILIACL YEENEDGEVV
     RVRDSYVAIA NACCAPRFPT LGGRVVNVAQ IIELVMTCIL YVVVSGNLMY NSFPGLPVSQ
     KSWSIIATAV LLPCAFLKNL KAVSKFSLLC TLAHFVINIL VIAYCLSRAR DWAWEKVKFY
     IDVKKFPISI GIIVFSYTSQ IFLPSLEGNM QQPSEFHCMM NWTHIAACVL KGLFALVAYL
     TWADETKEVI TDNLPGSIRA VVNIFLVAKA LLSYPLPFFA AVEVLEKSLF QEGSRAFFPA
     CYGGDGRLKS WGLTLRCALV VFTLLMAIYV PHFALLMGLT GSLTGAGLCF LLPSLFHLRL
     LWRKLLWHQV FFDVAIFVIG GICSVSGFVH SLEGLIEAYR TNAED
//