ID GLSN_MEDSA Reviewed; 2194 AA.
AC Q03460; Q40360;
DT 01-OCT-1993, integrated into UniProtKB/Swiss-Prot.
DT 01-OCT-1993, sequence version 1.
DT 13-SEP-2023, entry version 132.
DE RecName: Full=Glutamate synthase [NADH], amyloplastic;
DE EC=1.4.1.14;
DE AltName: Full=NADH-GOGAT;
DE Flags: Precursor;
OS Medicago sativa (Alfalfa).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC rosids; fabids; Fabales; Fabaceae; Papilionoideae; 50 kb inversion clade;
OC NPAAA clade; Hologalegina; IRL clade; Trifolieae; Medicago.
OX NCBI_TaxID=3879;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 102-114, FUNCTION, TISSUE
RP SPECIFICITY, AND INDUCTION.
RC STRAIN=cv. Saranac;
RX PubMed=8453303; DOI=10.1105/tpc.5.2.215;
RA Gregerson R.G., Miller S.S., Twary S.N., Gantt J.S., Vance C.P.;
RT "Molecular characterization of NADH-dependent glutamate synthase from
RT alfalfa nodules.";
RL Plant Cell 5:215-226(1993).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND TISSUE SPECIFICITY.
RC STRAIN=cv. Saranac; TISSUE=Seedling;
RX PubMed=7550373; DOI=10.1046/j.1365-313x.1995.08030345.x;
RA Vance C.P., Miller S.S., Gregerson R.G., Samac D.A., Robinson D.L.,
RA Gantt J.S.;
RT "Alfalfa NADH-dependent glutamate synthase: structure of the gene and
RT importance in symbiotic N2 fixation.";
RL Plant J. 8:345-358(1995).
RN [3]
RP FUNCTION, SUBUNIT, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP ACTIVITY REGULATION.
RC STRAIN=cv. Saranac;
RX PubMed=16666762; DOI=10.1104/pp.90.1.351;
RA Anderson M.P., Vance C.P., Heichel G.H., Miller S.S.;
RT "Purification and characterization of NADH-glutamate synthase from alfalfa
RT root nodules.";
RL Plant Physiol. 90:351-358(1989).
RN [4]
RP TISSUE SPECIFICITY.
RC STRAIN=cv. Saranac;
RX PubMed=10069821; DOI=10.1104/pp.119.3.817;
RA Trepp G.B., van de Mortel M., Yoshioka H., Miller S.S., Samac D.A.,
RA Gantt J.S., Vance C.P.;
RT "NADH-glutamate synthase in alfalfa root nodules. Genetic regulation and
RT cellular expression.";
RL Plant Physiol. 119:817-828(1999).
RN [5]
RP TISSUE SPECIFICITY, AND SUBCELLULAR LOCATION.
RC STRAIN=cv. Saranac;
RX PubMed=10069822; DOI=10.1104/pp.119.3.829;
RA Trepp G.B., Plank D.W., Stephen Gantt J., Vance C.P.;
RT "NADH-Glutamate synthase in alfalfa root nodules. Immunocytochemical
RT localization.";
RL Plant Physiol. 119:829-838(1999).
CC -!- FUNCTION: Required for the assimilation of symbiotically fixed nitrogen
CC into amino acids in root nodules. {ECO:0000269|PubMed:16666762,
CC ECO:0000269|PubMed:8453303}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2 L-glutamate + NAD(+) = 2-oxoglutarate + H(+) + L-glutamine +
CC NADH; Xref=Rhea:RHEA:13753, ChEBI:CHEBI:15378, ChEBI:CHEBI:16810,
CC ChEBI:CHEBI:29985, ChEBI:CHEBI:57540, ChEBI:CHEBI:57945,
CC ChEBI:CHEBI:58359; EC=1.4.1.14;
CC Evidence={ECO:0000269|PubMed:16666762};
CC -!- COFACTOR:
CC Name=[3Fe-4S] cluster; Xref=ChEBI:CHEBI:21137;
CC Note=Binds 1 [3Fe-4S] cluster.;
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC -!- COFACTOR:
CC Name=FMN; Xref=ChEBI:CHEBI:58210;
CC -!- ACTIVITY REGULATION: Inhibited by malate, citrate, glutamate, NAD(+)
CC and azaserine, but not by 2-2' dipyridil and N-ethylmaleimide.
CC {ECO:0000269|PubMed:16666762}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=466 uM for glutamine {ECO:0000269|PubMed:16666762};
CC KM=33 uM for 2-oxoglutarate {ECO:0000269|PubMed:16666762};
CC KM=4.2 uM for NADH {ECO:0000269|PubMed:16666762};
CC pH dependence:
CC Optimum pH is 7.5-8.5. {ECO:0000269|PubMed:16666762};
CC -!- PATHWAY: Amino-acid biosynthesis; L-glutamate biosynthesis via GLT
CC pathway; L-glutamate from 2-oxoglutarate and L-glutamine (NAD(+)
CC route): step 1/1.
CC -!- PATHWAY: Energy metabolism; nitrogen metabolism.
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:16666762}.
CC -!- SUBCELLULAR LOCATION: Plastid, amyloplast
CC {ECO:0000269|PubMed:10069822}.
CC -!- TISSUE SPECIFICITY: Expressed in infected cells in root nodules. Barely
CC detected in roots and stems. {ECO:0000269|PubMed:10069821,
CC ECO:0000269|PubMed:10069822, ECO:0000269|PubMed:7550373,
CC ECO:0000269|PubMed:8453303}.
CC -!- INDUCTION: Up-regulated during nodule development.
CC {ECO:0000269|PubMed:8453303}.
CC -!- SIMILARITY: Belongs to the glutamate synthase family. {ECO:0000305}.
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DR EMBL; L01660; AAB46617.1; -; mRNA.
DR EMBL; L37606; AAB41904.1; -; Genomic_DNA.
DR PIR; JQ1977; JQ1977.
DR AlphaFoldDB; Q03460; -.
DR SMR; Q03460; -.
DR UniPathway; UPA00045; -.
DR UniPathway; UPA00634; UER00690.
DR GO; GO:0009501; C:amyloplast; IEA:UniProtKB-SubCell.
DR GO; GO:0051538; F:3 iron, 4 sulfur cluster binding; IEA:UniProtKB-KW.
DR GO; GO:0050660; F:flavin adenine dinucleotide binding; IEA:InterPro.
DR GO; GO:0010181; F:FMN binding; IEA:InterPro.
DR GO; GO:0016040; F:glutamate synthase (NADH) activity; IEA:UniProtKB-EC.
DR GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR GO; GO:0006541; P:glutamine metabolic process; IEA:UniProtKB-KW.
DR GO; GO:0097054; P:L-glutamate biosynthetic process; IEA:UniProtKB-UniPathway.
DR CDD; cd00982; gltB_C; 1.
DR CDD; cd00713; GltS; 1.
DR CDD; cd02808; GltS_FMN; 1.
DR Gene3D; 3.20.20.70; Aldolase class I; 2.
DR Gene3D; 1.10.1060.10; Alpha-helical ferredoxin; 1.
DR Gene3D; 3.50.50.60; FAD/NAD(P)-binding domain; 1.
DR Gene3D; 2.160.20.60; Glutamate synthase, alpha subunit, C-terminal domain; 1.
DR Gene3D; 3.60.20.10; Glutamine Phosphoribosylpyrophosphate, subunit 1, domain 1; 1.
DR Gene3D; 3.40.50.720; NAD(P)-binding Rossmann-like Domain; 1.
DR InterPro; IPR013785; Aldolase_TIM.
DR InterPro; IPR028261; DPD_II.
DR InterPro; IPR036188; FAD/NAD-bd_sf.
DR InterPro; IPR023753; FAD/NAD-binding_dom.
DR InterPro; IPR017932; GATase_2_dom.
DR InterPro; IPR002489; Glu_synth_asu_C.
DR InterPro; IPR036485; Glu_synth_asu_C_sf.
DR InterPro; IPR006982; Glu_synth_centr_N.
DR InterPro; IPR012220; Glu_synth_euk.
DR InterPro; IPR002932; Glu_synthdom.
DR InterPro; IPR006005; Glut_synth_ssu1.
DR InterPro; IPR009051; Helical_ferredxn.
DR InterPro; IPR029055; Ntn_hydrolases_N.
DR NCBIfam; TIGR01317; GOGAT_sm_gam; 1.
DR PANTHER; PTHR43100; GLUTAMATE SYNTHASE [NADPH] SMALL CHAIN; 1.
DR PANTHER; PTHR43100:SF1; GLUTAMATE SYNTHASE [NADPH] SMALL CHAIN; 1.
DR Pfam; PF14691; Fer4_20; 1.
DR Pfam; PF00310; GATase_2; 1.
DR Pfam; PF04898; Glu_syn_central; 1.
DR Pfam; PF01645; Glu_synthase; 1.
DR Pfam; PF01493; GXGXG; 1.
DR Pfam; PF07992; Pyr_redox_2; 1.
DR PIRSF; PIRSF000187; GOGAT; 1.
DR PRINTS; PR00419; ADXRDTASE.
DR SUPFAM; SSF69336; Alpha subunit of glutamate synthase, C-terminal domain; 1.
DR SUPFAM; SSF46548; alpha-helical ferredoxin; 1.
DR SUPFAM; SSF51905; FAD/NAD(P)-binding domain; 1.
DR SUPFAM; SSF51395; FMN-linked oxidoreductases; 1.
DR SUPFAM; SSF56235; N-terminal nucleophile aminohydrolases (Ntn hydrolases); 1.
DR PROSITE; PS51278; GATASE_TYPE_2; 1.
PE 1: Evidence at protein level;
KW 3Fe-4S; Amino-acid biosynthesis; Amyloplast; Direct protein sequencing;
KW FAD; Flavoprotein; FMN; Glutamate biosynthesis; Glutamine amidotransferase;
KW Iron; Iron-sulfur; Metal-binding; NAD; Oxidoreductase; Plastid;
KW Transit peptide.
FT TRANSIT 1..101
FT /note="Amyloplast"
FT /evidence="ECO:0000269|PubMed:8453303"
FT CHAIN 102..2194
FT /note="Glutamate synthase [NADH], amyloplastic"
FT /id="PRO_0000011617"
FT DOMAIN 102..503
FT /note="Glutamine amidotransferase type-2"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00609"
FT REGION 1021..1045
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT ACT_SITE 102
FT /note="Nucleophile"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00609"
FT BINDING 1193..1250
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000250"
FT BINDING 1246
FT /ligand="[3Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:21137"
FT /evidence="ECO:0000250"
FT BINDING 1252
FT /ligand="[3Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:21137"
FT /evidence="ECO:0000250"
FT BINDING 1257
FT /ligand="[3Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:21137"
FT /evidence="ECO:0000250"
FT BINDING 1974..1988
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000255"
FT CONFLICT 33
FT /note="V -> F (in Ref. 2; AAB41904)"
FT /evidence="ECO:0000305"
FT CONFLICT 57
FT /note="L -> P (in Ref. 2; AAB41904)"
FT /evidence="ECO:0000305"
FT CONFLICT 185
FT /note="K -> M (in Ref. 2; AAB41904)"
FT /evidence="ECO:0000305"
FT CONFLICT 742
FT /note="H -> R (in Ref. 2; AAB41904)"
FT /evidence="ECO:0000305"
FT CONFLICT 989
FT /note="S -> G (in Ref. 2; AAB41904)"
FT /evidence="ECO:0000305"
FT CONFLICT 1503
FT /note="L -> Q (in Ref. 2; AAB41904)"
FT /evidence="ECO:0000305"
FT CONFLICT 2154
FT /note="A -> R (in Ref. 2; AAB41904)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 2194 AA; 240374 MW; 370A1B0F178367C4 CRC64;
MSNSLSLTFT ALNNPQINAI SNPNARLRPL ARVTRCSATC VERKRWLGTK LRSGGGLERI
QLWESGGLGR LPKLRVAVKS SFSAVPDKPM GLYDPAFDKD SCGVGFVAEL NGQSSRKTVT
DALEMLVRMT HRGACGCEAN TGDGAGILVA LPHGFYQEVV DFQLPPQGNY AVGMFFLPKS
DSRRKESKNI FTKVAESLGH KVLGWRSVPT DNTGLGKSAQ LTEPVIEQVF LTPSSDSKVD
LEKQMYILRK LSMVSITSAL NLQSDGITDF YICSLSSRTV IYKGQLTPAQ LGEYYYADLG
NERFTSYMAL IHSRFSTNTF PSWDRAQPFR VLGHNGEINT LRGNVNWIKA REGLLKCKEL
GLSENDLKKF LPIVDANSSD SGCFDGVLEF LLHSGKSLPE AVMMMIPEAW QNDKNMDPQR
KAFYEYYSAL MEPWDGPALI SFTDGHYLGA TLDRNGLRPG RFYVTHSGRV IMASEVGVVD
IPPEDVCRKG RLNPGMMLLV DFEKQIVVND DALKEQYSLA RPYGDWLEKQ KIELKDIIDS
VHESDIVPPT ISGVPPLSND DVDMENMGIQ GLLAPLKAFG YSVESLEILL LPMAKDGVEA
LGSMGNDTPL AVMSNREKLT FEYFKQMFAQ VTNPPIDPIR EKIVTSMRCM VGPEGDLTET
TEEQCHRLSL KGPLLSTKEM EAIKKMNYRG WRSKVIDITY SKERGTKGLE EALDRICTEA
HNAISEGYTT LVLSDRAFSK KHVAVSSLLA VGAVHQHLVK TLERTRVALM VESAEPREVH
HFCTLVGFGA DAICPYLAIE AIWRLQVDGK IPPKASGDFN SKDELVKKYF KASTYGMMKV
LAKMGISTLA SYKGAQIFEA LGLSSEVIEK CFAGTPSRVE GATFEMLAQD ALHLHELAFP
SRIFSPGSAE AVALPNPGDY HWRKGGEVHL NDPLAIAKLQ EAARTNSVDA YKQYSKTIHE
LNKACNLRGL LKFKDAASKV PISEVEPASE IVKRFCTGAM SYGSISLEAH TALATAMNTI
GGKSNTGEGG EQPSRMEPLA DGSRNPKRSA IKQVASGRFG VSSYYLTNAD ELQIKMAQGA
KPGEGGELPG HKVIGDIAIT RNSTAGVGLI SPPPHHDIYS IEDLAQLIHD LKNANPAARI
SVKLVSEAGV GVIASGVVKG HAEHVLISGH DGGTGASRWT GIKSAGLPWE LGLAETHQTL
VANDLRGRTT LQTDGQLKTG RDVAIAALLG AEEYGFSTAP LITLGCIMMR KCHKNTCPVG
IATQDPVLRE KFAGEPEHVI NFFFMVAEEM REIMSQLGFR TVNEMVGRSD MLEVDKEVVK
GNAKLENIDL SLLLRPAAEL RPEAAQYCVQ KQDHGLDMAL DNKLISLSNA ALEKGLPVYI
ETPICNTNRA VGTMLSHEVT KRYNLAGLPA DTIHIQFTGS AGQSFGAFLC PGITLELEGD
SNDYIGKGLS GGKVVVYPPK GSNFDPKDNI LIGNVALYGA TRGEAYFNGM AAERFCVRNS
GALAVVEGVG DHGCEYMTGG TVVVLGKTGR NFAAGMSGGI AYVLDVDGTF QSRCNLELVD
LDKVEEEEDI ITLRMLIQQH QRHTNSLLAK EVLVDFENLL PKFVKVFPRE YKRVLASMKS
DAASKDAVER AAEDVDEQDD EAQAVEKDAF EELKKLATAS LNEKPSEAPK RPSQVTDAVK
HRGFVAYERE GVQYRDPNVR LNDWNEVMME TKPGPLLKTQ SARCMDCGTP FCHQENSGCP
LGNKIPEFNE LVYQNRWQEA LERLLETNNF PEFTGRVCPA PCEGSCVLGI IENPVSIKNI
ECAIIDKAFE EGWMIPRPPV KRTGKRVAIV GSGPSGLAAA DQLNKMGHIV TVFERADRIG
GLMMYGVPNM KTDKVDIVQR RVNLMAEEGI NFVVNANIGL DPLYSLERLR EENDAIVLAV
GATKPRDLPV PGRELSGVHF AMEFLHANTK SLLDSNLQDG NYISAKGKKV VVIGGGDTGT
DCIGTSIRHG CTAVVNLELL PQPPPTRAPG NPWPQWPRIF RVDYGHQEAE TKFGKDPRTY
EVLTKRFVGD ENGVVKGLEV VRVCWEKDET GKFQFKEIEG SEEIIEADLV LLAMGFLGPE
ATIAEKLGVE RDNRSNFKAD YGRFSTSVDG VFAAGDCRRG QSLVVWAISE GRQAAAQVDS
YLTNEDHGID GNQDEFVKRQ QDLNKKHSKH TVMT
//
