NADA_ECOLI
ID NADA_ECOLI Reviewed; 347 AA.
AC P11458; P77373;
DT 01-OCT-1989, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1997, sequence version 3.
DT 03-AUG-2022, entry version 168.
DE RecName: Full=Quinolinate synthase {ECO:0000255|HAMAP-Rule:MF_00567, ECO:0000303|PubMed:18651751};
DE EC=2.5.1.72 {ECO:0000255|HAMAP-Rule:MF_00567, ECO:0000269|PubMed:10648170, ECO:0000269|PubMed:15898769, ECO:0000269|PubMed:15967443, ECO:0000269|PubMed:18674537};
DE AltName: Full=Quinolinate synthetase A {ECO:0000303|PubMed:2841129};
GN Name=nadA {ECO:0000255|HAMAP-Rule:MF_00567, ECO:0000303|PubMed:2841129};
GN Synonyms=nicA; OrderedLocusNames=b0750, JW0733;
OS Escherichia coli (strain K12).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=83333;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=2841129; DOI=10.1111/j.1432-1033.1988.tb14187.x;
RA Flachmann R., Kunz N., Seifert J., Guetlich M., Wientjes F.-J., Laeufer A.,
RA Gassen H.G.;
RT "Molecular biology of pyridine nucleotide biosynthesis in Escherichia coli.
RT Cloning and characterization of quinolinate synthesis genes nadA and
RT nadB.";
RL Eur. J. Biochem. 175:221-228(1988).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=8905232; DOI=10.1093/dnares/3.3.137;
RA Oshima T., Aiba H., Baba T., Fujita K., Hayashi K., Honjo A., Ikemoto K.,
RA Inada T., Itoh T., Kajihara M., Kanai K., Kashimoto K., Kimura S.,
RA Kitagawa M., Makino K., Masuda S., Miki T., Mizobuchi K., Mori H.,
RA Motomura K., Nakamura Y., Nashimoto H., Nishio Y., Saito N., Sampei G.,
RA Seki Y., Tagami H., Takemoto K., Wada C., Yamamoto Y., Yano M.,
RA Horiuchi T.;
RT "A 718-kb DNA sequence of the Escherichia coli K-12 genome corresponding to
RT the 12.7-28.0 min region on the linkage map.";
RL DNA Res. 3:137-155(1996).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / MG1655 / ATCC 47076;
RX PubMed=9278503; DOI=10.1126/science.277.5331.1453;
RA Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V.,
RA Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F.,
RA Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B.,
RA Shao Y.;
RT "The complete genome sequence of Escherichia coli K-12.";
RL Science 277:1453-1462(1997).
RN [4]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=16738553; DOI=10.1038/msb4100049;
RA Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S.,
RA Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.;
RT "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655
RT and W3110.";
RL Mol. Syst. Biol. 2:E1-E5(2006).
RN [5]
RP PROTEIN SEQUENCE OF 1-45, MASS SPECTROMETRY, FUNCTION, CATALYTIC ACTIVITY,
RP AND PATHWAY.
RX PubMed=10648170; DOI=10.1006/prep.1999.1153;
RA Ceciliani F., Caramori T., Ronchi S., Tedeschi G., Mortarino M.,
RA Galizzi A.;
RT "Cloning, overexpression, and purification of Escherichia coli quinolinate
RT synthetase.";
RL Protein Expr. Purif. 18:64-70(2000).
RN [6]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, AND SUBUNIT.
RC STRAIN=K12 / MG1655 / ATCC 47076;
RX PubMed=15967443; DOI=10.1016/j.febslet.2005.05.065;
RA Ollagnier-de Choudens S., Loiseau L., Sanakis Y., Barras F., Fontecave M.;
RT "Quinolinate synthetase, an iron-sulfur enzyme in NAD biosynthesis.";
RL FEBS Lett. 579:3737-3743(2005).
RN [7]
RP FUNCTION, CATALYTIC ACTIVITY, AND COFACTOR.
RX PubMed=15898769; DOI=10.1021/ja051369x;
RA Cicchillo R.M., Tu L., Stromberg J.A., Hoffart L.M., Krebs C., Booker S.J.;
RT "Escherichia coli quinolinate synthetase does indeed harbor a [4Fe-4S]
RT cluster.";
RL J. Am. Chem. Soc. 127:7310-7311(2005).
RN [8]
RP REGULATION BY DISULFIDE-BOND FORMATION, AND MUTAGENESIS OF CYS-291 AND
RP CYS-294.
RX PubMed=18651751; DOI=10.1021/bi801135y;
RA Saunders A.H., Booker S.J.;
RT "Regulation of the activity of Escherichia coli quinolinate synthase by
RT reversible disulfide-bond formation.";
RL Biochemistry 47:8467-8469(2008).
RN [9]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, SUBUNIT, DISULFIDE BOND, AND
RP MUTAGENESIS OF CYS-113; CYS-119; CYS-200; CYS-291; CYS-294 AND CYS-297.
RX PubMed=18674537; DOI=10.1016/j.febslet.2008.07.032;
RA Rousset C., Fontecave M., Ollagnier de Choudens S.;
RT "The [4Fe-4S] cluster of quinolinate synthase from Escherichia coli:
RT investigation of cluster ligands.";
RL FEBS Lett. 582:2937-2944(2008).
RN [10]
RP MUTAGENESIS OF TYR-49; HIS-203; GLU-228 AND THR-262.
RX PubMed=24650327; DOI=10.1021/ja501431b;
RA Cherrier M.V., Chan A., Darnault C., Reichmann D., Amara P.,
RA Ollagnier de Choudens S., Fontecilla-Camps J.C.;
RT "The crystal structure of FeS quinolinate synthase unravels an enzymatic
RT dehydration mechanism that uses tyrosine and a hydrolase-type triad.";
RL J. Am. Chem. Soc. 136:5253-5256(2014).
CC -!- FUNCTION: Catalyzes the condensation of iminoaspartate with
CC dihydroxyacetone phosphate to form quinolinate. {ECO:0000255|HAMAP-
CC Rule:MF_00567, ECO:0000269|PubMed:10648170,
CC ECO:0000269|PubMed:15898769, ECO:0000269|PubMed:15967443,
CC ECO:0000269|PubMed:18674537}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=dihydroxyacetone phosphate + iminosuccinate = H(+) + 2 H2O +
CC phosphate + quinolinate; Xref=Rhea:RHEA:25888, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:29959, ChEBI:CHEBI:43474,
CC ChEBI:CHEBI:57642, ChEBI:CHEBI:77875; EC=2.5.1.72;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_00567,
CC ECO:0000269|PubMed:10648170, ECO:0000269|PubMed:15898769,
CC ECO:0000269|PubMed:15967443, ECO:0000269|PubMed:18674537};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:25889;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_00567,
CC ECO:0000269|PubMed:10648170, ECO:0000269|PubMed:15898769,
CC ECO:0000269|PubMed:15967443, ECO:0000269|PubMed:18674537};
CC -!- COFACTOR:
CC Name=[4Fe-4S] cluster; Xref=ChEBI:CHEBI:49883;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_00567,
CC ECO:0000269|PubMed:15898769, ECO:0000269|PubMed:15967443,
CC ECO:0000269|PubMed:18674537};
CC Note=Binds 1 [4Fe-4S] cluster per subunit. {ECO:0000255|HAMAP-
CC Rule:MF_00567, ECO:0000269|PubMed:15898769,
CC ECO:0000269|PubMed:15967443, ECO:0000269|PubMed:18674537};
CC -!- ACTIVITY REGULATION: Activity is greater under oxic conditions and is
CC regulated by the formation of a reversible disulfide-bond between Cys-
CC 291 and Cys-294. {ECO:0000269|PubMed:18651751}.
CC -!- PATHWAY: Cofactor biosynthesis; NAD(+) biosynthesis; quinolinate from
CC iminoaspartate: step 1/1. {ECO:0000255|HAMAP-Rule:MF_00567,
CC ECO:0000269|PubMed:10648170}.
CC -!- SUBUNIT: Monomer (PubMed:18674537). Homodimer (PubMed:15967443).
CC {ECO:0000269|PubMed:15967443, ECO:0000269|PubMed:18674537}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000255|HAMAP-Rule:MF_00567,
CC ECO:0000305}.
CC -!- MASS SPECTROMETRY: Mass=38246; Method=MALDI;
CC Evidence={ECO:0000269|PubMed:10648170};
CC -!- SIMILARITY: Belongs to the quinolinate synthase family. Type 1
CC subfamily. {ECO:0000255|HAMAP-Rule:MF_00567, ECO:0000305}.
CC -!- SEQUENCE CAUTION:
CC Sequence=CAA31216.1; Type=Frameshift; Evidence={ECO:0000305};
CC ---------------------------------------------------------------------------
CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC ---------------------------------------------------------------------------
DR EMBL; X12713; CAA31216.1; ALT_FRAME; Genomic_DNA.
DR EMBL; U00096; AAC73837.1; -; Genomic_DNA.
DR EMBL; AP009048; BAA35409.1; -; Genomic_DNA.
DR PIR; F64810; SYECQA.
DR RefSeq; NP_415271.1; NC_000913.3.
DR RefSeq; WP_000115290.1; NZ_SSZK01000002.1.
DR AlphaFoldDB; P11458; -.
DR SMR; P11458; -.
DR BioGRID; 4263028; 14.
DR BioGRID; 849727; 2.
DR DIP; DIP-1027N; -.
DR IntAct; P11458; 6.
DR STRING; 511145.b0750; -.
DR jPOST; P11458; -.
DR PaxDb; P11458; -.
DR PRIDE; P11458; -.
DR EnsemblBacteria; AAC73837; AAC73837; b0750.
DR EnsemblBacteria; BAA35409; BAA35409; BAA35409.
DR GeneID; 945351; -.
DR KEGG; ecj:JW0733; -.
DR KEGG; eco:b0750; -.
DR PATRIC; fig|1411691.4.peg.1529; -.
DR EchoBASE; EB0624; -.
DR eggNOG; COG0379; Bacteria.
DR HOGENOM; CLU_047382_1_0_6; -.
DR InParanoid; P11458; -.
DR OMA; CFCSTMN; -.
DR PhylomeDB; P11458; -.
DR BioCyc; EcoCyc:QUINOLINATE-SYNTHA-MON; -.
DR BioCyc; MetaCyc:QUINOLINATE-SYNTHA-MON; -.
DR BRENDA; 2.5.1.72; 2026.
DR UniPathway; UPA00253; UER00327.
DR PRO; PR:P11458; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005829; C:cytosol; IDA:EcoCyc.
DR GO; GO:0051539; F:4 iron, 4 sulfur cluster binding; IDA:EcoCyc.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0008987; F:quinolinate synthetase A activity; IDA:EcoCyc.
DR GO; GO:0034628; P:'de novo' NAD biosynthetic process from aspartate; IMP:EcoCyc.
DR Gene3D; 3.40.50.10800; -; 3.
DR HAMAP; MF_00567; NadA_type1; 1.
DR InterPro; IPR003473; NadA.
DR InterPro; IPR036094; NadA_sf.
DR InterPro; IPR023513; Quinolinate_synth_A_type1.
DR PANTHER; PTHR30573; PTHR30573; 1.
DR Pfam; PF02445; NadA; 1.
DR SUPFAM; SSF142754; SSF142754; 1.
DR TIGRFAMs; TIGR00550; nadA; 1.
PE 1: Evidence at protein level;
KW 4Fe-4S; Cytoplasm; Direct protein sequencing; Disulfide bond; Iron;
KW Iron-sulfur; Metal-binding; Pyridine nucleotide biosynthesis;
KW Reference proteome; Transferase.
FT CHAIN 1..347
FT /note="Quinolinate synthase"
FT /id="PRO_0000155760"
FT BINDING 47
FT /ligand="iminosuccinate"
FT /ligand_id="ChEBI:CHEBI:77875"
FT /evidence="ECO:0000250|UniProtKB:O57767, ECO:0000255|HAMAP-
FT Rule:MF_00567"
FT BINDING 68
FT /ligand="iminosuccinate"
FT /ligand_id="ChEBI:CHEBI:77875"
FT /evidence="ECO:0000250|UniProtKB:O57767, ECO:0000255|HAMAP-
FT Rule:MF_00567"
FT BINDING 113
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00567,
FT ECO:0000269|PubMed:18674537"
FT BINDING 139..141
FT /ligand="iminosuccinate"
FT /ligand_id="ChEBI:CHEBI:77875"
FT /evidence="ECO:0000250|UniProtKB:O57767, ECO:0000255|HAMAP-
FT Rule:MF_00567"
FT BINDING 156
FT /ligand="iminosuccinate"
FT /ligand_id="ChEBI:CHEBI:77875"
FT /evidence="ECO:0000250|UniProtKB:O57767, ECO:0000255|HAMAP-
FT Rule:MF_00567"
FT BINDING 200
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00567,
FT ECO:0000269|PubMed:18674537"
FT BINDING 226..228
FT /ligand="iminosuccinate"
FT /ligand_id="ChEBI:CHEBI:77875"
FT /evidence="ECO:0000250|UniProtKB:O57767, ECO:0000255|HAMAP-
FT Rule:MF_00567"
FT BINDING 243
FT /ligand="iminosuccinate"
FT /ligand_id="ChEBI:CHEBI:77875"
FT /evidence="ECO:0000250|UniProtKB:O57767, ECO:0000255|HAMAP-
FT Rule:MF_00567"
FT BINDING 297
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00567,
FT ECO:0000269|PubMed:18674537"
FT DISULFID 291..294
FT /evidence="ECO:0000269|PubMed:18651751,
FT ECO:0000269|PubMed:18674537"
FT MUTAGEN 49
FT /note="Y->F: 13-fold decrease in specific activity in the
FT presence of NadB."
FT /evidence="ECO:0000269|PubMed:24650327"
FT MUTAGEN 113
FT /note="C->A: Loss of activity. Strong decrease in iron
FT content. Forms oligomers."
FT /evidence="ECO:0000269|PubMed:18674537"
FT MUTAGEN 119
FT /note="C->A: Retains 30% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:18674537"
FT MUTAGEN 200
FT /note="C->A: Loss of activity. Strong decrease in iron
FT content. Forms oligomers."
FT /evidence="ECO:0000269|PubMed:18674537"
FT MUTAGEN 203
FT /note="H->F: Loss of activity."
FT /evidence="ECO:0000269|PubMed:24650327"
FT MUTAGEN 228
FT /note="E->A: 20-fold decrease in specific activity in the
FT presence of NadB."
FT /evidence="ECO:0000269|PubMed:24650327"
FT MUTAGEN 262
FT /note="T->V: 12-fold decrease in specific activity in the
FT presence of NadB."
FT /evidence="ECO:0000269|PubMed:24650327"
FT MUTAGEN 291
FT /note="C->A: Retains 5% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:18674537"
FT MUTAGEN 291
FT /note="C->S: Activity is 3.3-fold lower under oxic
FT conditions than under anoxic conditions."
FT /evidence="ECO:0000269|PubMed:18651751"
FT MUTAGEN 294
FT /note="C->A: Retains 3% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:18674537"
FT MUTAGEN 294
FT /note="C->S: Activity is 13-fold lower under oxic
FT conditions than under anoxic conditions."
FT /evidence="ECO:0000269|PubMed:18651751"
FT MUTAGEN 297
FT /note="C->A: Loss of activity. Strong decrease in iron
FT content. Forms oligomers."
FT /evidence="ECO:0000269|PubMed:18674537"
FT CONFLICT 35..36
FT /note="RL -> LR (in Ref. 1; CAA31216)"
FT /evidence="ECO:0000305"
FT CONFLICT 153
FT /note="V -> G (in Ref. 1; CAA31216)"
FT /evidence="ECO:0000305"
FT CONFLICT 170
FT /note="G -> C (in Ref. 1; CAA31216)"
FT /evidence="ECO:0000305"
FT CONFLICT 329
FT /note="E -> R (in Ref. 1; CAA31216)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 347 AA; 38241 MW; 17014F7621E5EC6B CRC64;
MSVMFDPDTA IYPFPPKPTP LSIDEKAYYR EKIKRLLKER NAVMVAHYYT DPEIQQLAEE
TGGCISDSLE MARFGAKHPA STLLVAGVRF MGETAKILSP EKTILMPTLQ AECSLDLGCP
VEEFNAFCDA HPDRTVVVYA NTSAAVKARA DWVVTSSIAV ELIDHLDSLG EKIIWAPDKH
LGRYVQKQTG GDILCWQGAC IVHDEFKTQA LTRLQEEYPD AAILVHPESP QAIVDMADAV
GSTSQLIAAA KTLPHQRLIV ATDRGIFYKM QQAVPDKELL EAPTAGEGAT CRSCAHCPWM
AMNGLQAIAE ALEQEGSNHE VHVDERLRER ALVPLNRMLD FAATLRG
