ACNA_ECOLI
ID ACNA_ECOLI Reviewed; 891 AA.
AC P25516; P78060; P78148;
DT 01-MAY-1992, integrated into UniProtKB/Swiss-Prot.
DT 15-JAN-2008, sequence version 3.
DT 03-AUG-2022, entry version 176.
DE RecName: Full=Aconitate hydratase A {ECO:0000303|PubMed:10585860};
DE Short=ACN {ECO:0000303|PubMed:10585860};
DE Short=Aconitase {ECO:0000303|PubMed:10585860};
DE EC=4.2.1.3 {ECO:0000269|PubMed:10585860};
DE AltName: Full=Iron-responsive protein-like {ECO:0000305|PubMed:10589714};
DE Short=IRP-like {ECO:0000305|PubMed:10589714};
DE AltName: Full=RNA-binding protein {ECO:0000303|PubMed:10589714};
DE AltName: Full=Stationary phase enzyme {ECO:0000303|PubMed:10589714};
GN Name=acnA {ECO:0000303|PubMed:9421904}; Synonyms=acn;
GN OrderedLocusNames=b1276, JW1268;
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=1541275; DOI=10.1111/j.1432-1033.1992.tb16673.x;
RA Prodromou C., Artymiuk P.J., Guest J.R.;
RT "The aconitase of Escherichia coli. Nucleotide sequence of the aconitase
RT gene and amino acid sequence similarity with mitochondrial aconitases, the
RT iron-responsive-element-binding protein and isopropylmalate isomerases.";
RL Eur. J. Biochem. 204:599-609(1992).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=9097039; DOI=10.1093/dnares/3.6.363;
RA Aiba H., Baba T., Fujita K., Hayashi K., Inada T., Isono K., Itoh T.,
RA Kasai H., Kashimoto K., Kimura S., Kitakawa M., Kitagawa M., Makino K.,
RA Miki T., Mizobuchi K., Mori H., Mori T., Motomura K., Nakade S.,
RA Nakamura Y., Nashimoto H., Nishio Y., Oshima T., Saito N., Sampei G.,
RA Seki Y., Sivasundaram S., Tagami H., Takeda J., Takemoto K., Takeuchi Y.,
RA Wada C., Yamamoto Y., Horiuchi T.;
RT "A 570-kb DNA sequence of the Escherichia coli K-12 genome corresponding to
RT the 28.0-40.1 min region on the linkage map.";
RL DNA Res. 3:363-377(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 2-19, FUNCTION, AND SUBUNIT.
RX PubMed=1838390; DOI=10.1099/00221287-137-11-2505;
RA Prodromou C., Haynes M.J., Guest J.R.;
RT "The aconitase of Escherichia coli: purification of the enzyme and
RT molecular cloning and map location of the gene (acn).";
RL J. Gen. Microbiol. 137:2505-2515(1991).
RN [6]
RP INDUCTION, AND DISRUPTION PHENOTYPE.
RX PubMed=8000525; DOI=10.1099/00221287-140-10-2531;
RA Gruer M.J., Guest J.R.;
RT "Two genetically-distinct and differentially-regulated aconitases (AcnA and
RT AcnB) in Escherichia coli.";
RL Microbiology 140:2531-2541(1994).
RN [7]
RP COFACTOR.
RX PubMed=7588761; DOI=10.1111/j.1432-1033.1995.317_1.x;
RA Bennett B., Gruer M.J., Guest J.R., Thomson A.J.;
RT "Spectroscopic characterisation of an aconitase (AcnA) of Escherichia
RT coli.";
RL Eur. J. Biochem. 233:317-326(1995).
RN [8]
RP DISRUPTION PHENOTYPE.
RX PubMed=9202458; DOI=10.1099/00221287-143-6-1837;
RA Gruer M.J., Bradbury A.J., Guest J.R.;
RT "Construction and properties of aconitase mutants of Escherichia coli.";
RL Microbiology 143:1837-1846(1997).
RN [9]
RP FUNCTION, AND INDUCTION.
RX PubMed=9421904; DOI=10.1099/00221287-143-12-3795;
RA Cunningham L., Gruer M.J., Guest J.R.;
RT "Transcriptional regulation of the aconitase genes (acnA and acnB) of
RT Escherichia coli.";
RL Microbiology 143:3795-3805(1997).
RN [10]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, MAGNETIC
RP CIRCULAR DICHROISM, EPR SPECTROSCOPY, AND COFACTOR.
RX PubMed=10585860; DOI=10.1042/bj3440739;
RA Jordan P.A., Tang Y., Bradbury A.J., Thomson A.J., Guest J.R.;
RT "Biochemical and spectroscopic characterization of Escherichia coli
RT aconitases (AcnA and AcnB).";
RL Biochem. J. 344:739-746(1999).
RN [11]
RP FUNCTION AS A RNA-BINDING PROTEIN.
RX PubMed=10589714; DOI=10.1099/00221287-145-11-3069;
RA Tang Y., Guest J.R.;
RT "Direct evidence for mRNA binding and post-transcriptional regulation by
RT Escherichia coli aconitases.";
RL Microbiology 145:3069-3079(1999).
RN [12]
RP FUNCTION, AND DISRUPTION PHENOTYPE.
RX PubMed=11932448; DOI=10.1099/00221287-148-4-1027;
RA Tang Y., Quail M.A., Artymiuk P.J., Guest J.R., Green J.;
RT "Escherichia coli aconitases and oxidative stress: post-transcriptional
RT regulation of sodA expression.";
RL Microbiology 148:1027-1037(2002).
RN [13]
RP INDUCTION.
RX PubMed=11917098; DOI=10.1073/pnas.032066599;
RA Masse E., Gottesman S.;
RT "A small RNA regulates the expression of genes involved in iron metabolism
RT in Escherichia coli.";
RL Proc. Natl. Acad. Sci. U.S.A. 99:4620-4625(2002).
RN [14]
RP FUNCTION, AND DISRUPTION PHENOTYPE.
RX PubMed=12486059; DOI=10.1128/jb.185.1.221-230.2003;
RA Varghese S., Tang Y., Imlay J.A.;
RT "Contrasting sensitivities of Escherichia coli aconitases A and B to
RT oxidation and iron depletion.";
RL J. Bacteriol. 185:221-230(2003).
RN [15]
RP INDUCTION.
RX PubMed=15672380; DOI=10.1002/bit.20389;
RA Lu C., Albano C.R., Bentley W.E., Rao G.;
RT "Quantitative and kinetic study of oxidative stress regulons using green
RT fluorescent protein.";
RL Biotechnol. Bioeng. 89:574-587(2005).
RN [16]
RP INDUCTION.
RX PubMed=17015655; DOI=10.1128/jb.00508-06;
RA Weber A., Kogl S.A., Jung K.;
RT "Time-dependent proteome alterations under osmotic stress during aerobic
RT and anaerobic growth in Escherichia coli.";
RL J. Bacteriol. 188:7165-7175(2006).
CC -!- FUNCTION: Catalyzes the reversible isomerization of citrate to
CC isocitrate via cis-aconitate. The apo form of AcnA functions as a RNA-
CC binding regulatory protein which plays a role as a maintenance or
CC survival enzyme during nutritional or oxidative stress. During
CC oxidative stress inactive AcnA apo-enzyme without iron sulfur clusters
CC binds the acnA mRNA 3' UTRs (untranslated regions), stabilizes acnA
CC mRNA and increases AcnA synthesis, thus mediating a post-
CC transcriptional positive autoregulatory switch. AcnA also enhances the
CC stability of the sodA transcript. {ECO:0000269|PubMed:10585860,
CC ECO:0000269|PubMed:10589714, ECO:0000269|PubMed:11932448,
CC ECO:0000269|PubMed:12486059, ECO:0000269|PubMed:1838390,
CC ECO:0000269|PubMed:9421904}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=citrate = D-threo-isocitrate; Xref=Rhea:RHEA:10336,
CC ChEBI:CHEBI:15562, ChEBI:CHEBI:16947; EC=4.2.1.3;
CC Evidence={ECO:0000269|PubMed:10585860};
CC -!- COFACTOR:
CC Name=[4Fe-4S] cluster; Xref=ChEBI:CHEBI:49883;
CC Evidence={ECO:0000269|PubMed:10585860, ECO:0000269|PubMed:7588761};
CC Note=Binds 1 [4Fe-4S] cluster per subunit.
CC {ECO:0000269|PubMed:10585860, ECO:0000269|PubMed:7588761};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=1.16 mM for citrate {ECO:0000269|PubMed:10585860};
CC KM=0.058 mM for cis-aconitate {ECO:0000269|PubMed:10585860};
CC KM=0.014 mM for isocitrate (using 0.01-0.8 mM substrate)
CC {ECO:0000269|PubMed:10585860};
CC KM=1.77 mM for isocitrate (using 0.8-40 mM substrate)
CC {ECO:0000269|PubMed:10585860};
CC Vmax=6.13 umol/min/mg enzyme with citrate as substrate
CC {ECO:0000269|PubMed:10585860};
CC Vmax=14.5 umol/min/mg enzyme with cis-aconitate as substrate
CC {ECO:0000269|PubMed:10585860};
CC Vmax=3.57 umol/min/mg enzyme using 0.01-0.8 mM isocitrate as
CC substrate {ECO:0000269|PubMed:10585860};
CC Vmax=14.7 umol/min/mg enzyme using 0.8-40 mM isocitrate as substrate
CC {ECO:0000269|PubMed:10585860};
CC pH dependence:
CC Optimum pH is 7.4. It retains a high specific activity over a broad
CC pH range. {ECO:0000269|PubMed:10585860};
CC -!- PATHWAY: Carbohydrate metabolism; tricarboxylic acid cycle; isocitrate
CC from oxaloacetate: step 2/2. {ECO:0000305}.
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:1838390}.
CC -!- INDUCTION: Induced upon entry into stationary phase by iron, oxidative
CC and salt stress under aerobic conditions. AcnA is subject to CRP-
CC mediated catabolite repression and ArcA-mediated anaerobic repression.
CC AcnA is negatively regulated by ryhB RNA. {ECO:0000269|PubMed:11917098,
CC ECO:0000269|PubMed:15672380, ECO:0000269|PubMed:17015655,
CC ECO:0000269|PubMed:8000525, ECO:0000269|PubMed:9421904}.
CC -!- DISRUPTION PHENOTYPE: Cells lacking this gene are more sensitive to
CC peroxide stress. The acnAB double mutant does not grow on
CC unsupplemented glucose minimal medium and does not respond under
CC aerobic conditions to glutamate. The acnAB double mutant retains a low
CC but significant aconitase activity. {ECO:0000269|PubMed:11932448,
CC ECO:0000269|PubMed:12486059, ECO:0000269|PubMed:8000525,
CC ECO:0000269|PubMed:9202458}.
CC -!- MISCELLANEOUS: The AcnA activity over a broad pH range might be a
CC useful adaptation for specifically expression in the stationary phase,
CC where the intracellular pH may vary over a wider range. AcnA is
CC resistant to oxidation in vivo. {ECO:0000269|PubMed:10585860,
CC ECO:0000269|PubMed:12486059}.
CC -!- SIMILARITY: Belongs to the aconitase/IPM isomerase family.
CC {ECO:0000305}.
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DR EMBL; X60293; CAA42834.1; -; Genomic_DNA.
DR EMBL; U00096; AAC74358.1; -; Genomic_DNA.
DR EMBL; AP009048; BAA14828.1; -; Genomic_DNA.
DR PIR; G64875; G64875.
DR RefSeq; NP_415792.1; NC_000913.3.
DR RefSeq; WP_000099535.1; NZ_LN832404.1.
DR AlphaFoldDB; P25516; -.
DR SMR; P25516; -.
DR BioGRID; 4260134; 23.
DR BioGRID; 851065; 2.
DR DIP; DIP-9043N; -.
DR IntAct; P25516; 13.
DR STRING; 511145.b1276; -.
DR jPOST; P25516; -.
DR PaxDb; P25516; -.
DR PRIDE; P25516; -.
DR EnsemblBacteria; AAC74358; AAC74358; b1276.
DR EnsemblBacteria; BAA14828; BAA14828; BAA14828.
DR GeneID; 946724; -.
DR KEGG; ecj:JW1268; -.
DR KEGG; eco:b1276; -.
DR PATRIC; fig|511145.12.peg.1327; -.
DR EchoBASE; EB1301; -.
DR eggNOG; COG1048; Bacteria.
DR HOGENOM; CLU_013476_2_1_6; -.
DR InParanoid; P25516; -.
DR PhylomeDB; P25516; -.
DR BioCyc; EcoCyc:ACONITASE-MON; -.
DR BioCyc; MetaCyc:ACONITASE-MON; -.
DR BRENDA; 4.2.1.3; 2026.
DR SABIO-RK; P25516; -.
DR UniPathway; UPA00223; UER00718.
DR PRO; PR:P25516; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IDA:EcoliWiki.
DR GO; GO:0005829; C:cytosol; IDA:EcoCyc.
DR GO; GO:0051539; F:4 iron, 4 sulfur cluster binding; IDA:EcoliWiki.
DR GO; GO:0003994; F:aconitate hydratase activity; IDA:EcoliWiki.
DR GO; GO:0047780; F:citrate dehydratase activity; IEA:UniProtKB-EC.
DR GO; GO:0005506; F:iron ion binding; IMP:EcoliWiki.
DR GO; GO:0030350; F:iron-responsive element binding; IBA:GO_Central.
DR GO; GO:0046872; F:metal ion binding; IDA:EcoliWiki.
DR GO; GO:0003730; F:mRNA 3'-UTR binding; IDA:EcoCyc.
DR GO; GO:0003729; F:mRNA binding; IDA:EcoliWiki.
DR GO; GO:0009061; P:anaerobic respiration; IDA:EcoliWiki.
DR GO; GO:0006101; P:citrate metabolic process; IBA:GO_Central.
DR GO; GO:0006097; P:glyoxylate cycle; NAS:EcoliWiki.
DR GO; GO:0006979; P:response to oxidative stress; IDA:EcoliWiki.
DR GO; GO:0006099; P:tricarboxylic acid cycle; IBA:GO_Central.
DR CDD; cd01580; AcnA_IRP_Swivel; 1.
DR Gene3D; 3.20.19.10; -; 1.
DR Gene3D; 3.30.499.10; -; 2.
DR InterPro; IPR044137; AcnA_IRP_Swivel.
DR InterPro; IPR015931; Acnase/IPM_dHydase_lsu_aba_1/3.
DR InterPro; IPR001030; Acoase/IPM_deHydtase_lsu_aba.
DR InterPro; IPR015928; Aconitase/3IPM_dehydase_swvl.
DR InterPro; IPR006249; Aconitase/IRP2.
DR InterPro; IPR018136; Aconitase_4Fe-4S_BS.
DR InterPro; IPR036008; Aconitase_4Fe-4S_dom.
DR InterPro; IPR000573; AconitaseA/IPMdHydase_ssu_swvl.
DR PANTHER; PTHR11670; PTHR11670; 1.
DR Pfam; PF00330; Aconitase; 1.
DR Pfam; PF00694; Aconitase_C; 1.
DR PRINTS; PR00415; ACONITASE.
DR SUPFAM; SSF53732; SSF53732; 1.
DR TIGRFAMs; TIGR01341; aconitase_1; 1.
DR PROSITE; PS00450; ACONITASE_1; 1.
DR PROSITE; PS01244; ACONITASE_2; 1.
PE 1: Evidence at protein level;
KW 4Fe-4S; Direct protein sequencing; Iron; Iron-sulfur; Lyase; Metal-binding;
KW Reference proteome; RNA-binding; Tricarboxylic acid cycle.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:1838390"
FT CHAIN 2..891
FT /note="Aconitate hydratase A"
FT /id="PRO_0000076661"
FT BINDING 435
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000250|UniProtKB:P36683"
FT BINDING 501
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000250|UniProtKB:P36683"
FT BINDING 504
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000250|UniProtKB:P36683"
FT CONFLICT 522
FT /note="S -> G (in Ref. 1; CAA42834 and 4; BAA14828)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 891 AA; 97677 MW; EB47E5B3C3F9C56C CRC64;
MSSTLREASK DTLQAKDKTY HYYSLPLAAK SLGDITRLPK SLKVLLENLL RWQDGNSVTE
EDIHALAGWL KNAHADREIA YRPARVLMQD FTGVPAVVDL AAMREAVKRL GGDTAKVNPL
SPVDLVIDHS VTVDRFGDDE AFEENVRLEM ERNHERYVFL KWGKQAFSRF SVVPPGTGIC
HQVNLEYLGK AVWSELQDGE WIAYPDTLVG TDSHTTMING LGVLGWGVGG IEAEAAMLGQ
PVSMLIPDVV GFKLTGKLRE GITATDLVLT VTQMLRKHGV VGKFVEFYGD GLDSLPLADR
ATIANMSPEY GATCGFFPID AVTLDYMRLS GRSEDQVELV EKYAKAQGMW RNPGDEPIFT
STLELDMNDV EASLAGPKRP QDRVALPDVP KAFAASNELE VNATHKDRQP VDYVMNGHQY
QLPDGAVVIA AITSCTNTSN PSVLMAAGLL AKKAVTLGLK RQPWVKASLA PGSKVVSDYL
AKAKLTPYLD ELGFNLVGYG CTTCIGNSGP LPDPIETAIK KSDLTVGAVL SGNRNFEGRI
HPLVKTNWLA SPPLVVAYAL AGNMNINLAS EPIGHDRKGD PVYLKDIWPS AQEIARAVEQ
VSTEMFRKEY AEVFEGTAEW KGINVTRSDT YGWQEDSTYI RLSPFFDEMQ ATPAPVEDIH
GARILAMLGD SVTTDHISPA GSIKPDSPAG RYLQGRGVER KDFNSYGSRR GNHEVMMRGT
FANIRIRNEM VPGVEGGMTR HLPDSDVVSI YDAAMRYKQE QTPLAVIAGK EYGSGSSRDW
AAKGPRLLGI RVVIAESFER IHRSNLIGMG ILPLEFPQGV TRKTLGLTGE EKIDIGDLQN
LQPGATVPVT LTRADGSQEV VPCRCRIDTA TELTYYQNDG ILHYVIRNML K
