ID   HEM1_ECOLI              Reviewed;         418 AA.
AC   P0A6X1; P13580; Q59405;
DT   29-MAR-2005, integrated into UniProtKB/Swiss-Prot.
DT   29-MAR-2005, sequence version 1.
DT   24-JAN-2024, entry version 141.
DE   RecName: Full=Glutamyl-tRNA reductase;
DE            Short=GluTR;
DE            EC=1.2.1.70;
GN   Name=hemA; OrderedLocusNames=b1210, JW1201;
OS   Escherichia coli (strain K12).
OC   Bacteria; Pseudomonadota; Gammaproteobacteria; Enterobacterales;
OC   Enterobacteriaceae; Escherichia.
OX   NCBI_TaxID=83333;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC   STRAIN=K12;
RX   PubMed=2684779; DOI=10.1016/0378-1119(89)90046-2;
RA   Li J.-M., Russell C.S., Cosloy S.D.;
RT   "Cloning and structure of the hem A gene of Escherichia coli K-12.";
RL   Gene 82:209-217(1989).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC   STRAIN=K12;
RX   PubMed=2548996; DOI=10.1128/jb.171.9.4728-4735.1989;
RA   Verkamp E., Chelm B.K.;
RT   "Isolation, nucleotide sequence, and preliminary characterization of the
RT   Escherichia coli K-12 hemA gene.";
RL   J. Bacteriol. 171:4728-4735(1989).
RN   [3]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC   STRAIN=K12;
RX   PubMed=2664455; DOI=10.1007/bf00334375;
RA   Drolet M., Peloquin L., Echelard Y., Cousineau L., Sasarman A.;
RT   "Isolation and nucleotide sequence of the hemA gene of Escherichia coli
RT   K12.";
RL   Mol. Gen. Genet. 216:347-352(1989).
RN   [4]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC   STRAIN=K12;
RX   PubMed=7543480; DOI=10.1128/jb.177.15.4488-4500.1995;
RA   Strohmaier H., Remler P., Renner W., Hoegenauer G.;
RT   "Expression of genes kdsA and kdsB involved in 3-deoxy-D-manno-octulosonic
RT   acid metabolism and biosynthesis of enterobacterial lipopolysaccharide is
RT   growth phase regulated primarily at the transcriptional level in
RT   Escherichia coli K-12.";
RL   J. Bacteriol. 177:4488-4500(1995).
RN   [5]
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   [6]
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   [7]
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   [8]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-49.
RX   PubMed=1427085; DOI=10.1016/0378-1119(92)90170-t;
RA   Ikemi M., Murakami K., Hashimoto M., Murooka Y.;
RT   "Cloning and characterization of genes involved in the biosynthesis of
RT   delta-aminolevulinic acid in Escherichia coli.";
RL   Gene 121:127-132(1992).
RN   [9]
RP   PROTEIN SEQUENCE OF 1-15, FUNCTION, CATALYTIC ACTIVITY, CHARACTERIZATION,
RP   ACTIVITY REGULATION, IDENTIFICATION BY MASS SPECTROMETRY, SUBUNIT, KINETIC
RP   PARAMETERS, CATALYTIC MECHANISM, AND MUTAGENESIS OF GLY-7; CYS-50; CYS-74;
RP   GLY-106; GLU-114; SER-145; CYS-170; GLY-191 AND ARG-314.
RX   PubMed=12370189; DOI=10.1074/jbc.m206924200;
RA   Schauer S., Chaturvedi S., Randau L., Moser J., Kitabatake M., Lorenz S.,
RA   Verkamp E., Schubert W.-D., Nakayashiki T., Murai M., Wall K.,
RA   Thomann H.-U., Heinz D.W., Inokuchi H., Soell D., Jahn D.;
RT   "Escherichia coli glutamyl-tRNA reductase. Trapping the thioester
RT   intermediate.";
RL   J. Biol. Chem. 277:48657-48663(2002).
RN   [10]
RP   FUNCTION.
RX   PubMed=1569081; DOI=10.1016/s0021-9258(18)42438-6;
RA   Verkamp E., Jahn M., Jahn D., Kumar A.M., Soell D.;
RT   "Glutamyl-tRNA reductase from Escherichia coli and Synechocystis 6803. Gene
RT   structure and expression.";
RL   J. Biol. Chem. 267:8275-8280(1992).
RN   [11]
RP   INTERACTION WITH GSA-AM.
RX   PubMed=15757895; DOI=10.1074/jbc.m500440200;
RA   Lueer C., Schauer S., Moebius K., Schulze J., Schubert W.-D., Heinz D.W.,
RA   Jahn D., Moser J.;
RT   "Complex formation between glutamyl-tRNA reductase and glutamate-1-
RT   semialdehyde 2,1-aminomutase in Escherichia coli during the initial
RT   reactions of porphyrin biosynthesis.";
RL   J. Biol. Chem. 280:18568-18572(2005).
RN   [12]
RP   MUTAGENESIS OF THR-49; ARG-52; GLU-54; HIS-99; SER-109 AND GLN-116.
RX   PubMed=17697121; DOI=10.1111/j.1742-4658.2007.05989.x;
RA   Lueer C., Schauer S., Virus S., Schubert W.-D., Heinz D.W., Moser J.,
RA   Jahn D.;
RT   "Glutamate recognition and hydride transfer by Escherichia coli glutamyl-
RT   tRNA reductase.";
RL   FEBS J. 274:4609-4614(2007).
CC   -!- FUNCTION: Catalyzes the NADPH-dependent reduction of glutamyl-tRNA(Glu)
CC       to glutamate 1-semialdehyde (GSA). In the absence of NADPH, exhibits
CC       substrate esterase activity, leading to the release of glutamate from
CC       tRNA. {ECO:0000269|PubMed:12370189, ECO:0000269|PubMed:1569081}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=(S)-4-amino-5-oxopentanoate + NADP(+) + tRNA(Glu) = H(+) + L-
CC         glutamyl-tRNA(Glu) + NADPH; Xref=Rhea:RHEA:12344, Rhea:RHEA-
CC         COMP:9663, Rhea:RHEA-COMP:9680, ChEBI:CHEBI:15378, ChEBI:CHEBI:57501,
CC         ChEBI:CHEBI:57783, ChEBI:CHEBI:58349, ChEBI:CHEBI:78442,
CC         ChEBI:CHEBI:78520; EC=1.2.1.70;
CC         Evidence={ECO:0000269|PubMed:12370189};
CC   -!- ACTIVITY REGULATION: Activated by Mg(2+) ions. Inhibited by metal-
CC       chelating agents such as EDTA, EGTA, 1,10-phenanthroline, 2,2'-
CC       dipyridyl, and by PtCl4 and KPdCl4 as well as Ni(2+) and Co(2+). Also
CC       inhibited by iodoacetamide, N-tosyl-L-phenylalanine chloromethyl
CC       ketone, and 5,5'-dithiobis(2-nitrobenzoic acid), as well as glutamycin.
CC       {ECO:0000269|PubMed:12370189}.
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=24 uM for L-glutamyl-tRNA(Glu) {ECO:0000269|PubMed:12370189};
CC         KM=39 uM for NADPH {ECO:0000269|PubMed:12370189};
CC   -!- PATHWAY: Porphyrin-containing compound metabolism; protoporphyrin-IX
CC       biosynthesis; 5-aminolevulinate from L-glutamyl-tRNA(Glu): step 1/2.
CC   -!- SUBUNIT: Homodimer. Interacts with glutamate-1-semialdehyde 2,1-
CC       aminomutase (GSA-AM), which forms a metabolic channeling between both
CC       enzymes to protect the reactive aldehyde species GSA.
CC       {ECO:0000269|PubMed:12370189, ECO:0000269|PubMed:15757895}.
CC   -!- DOMAIN: Possesses an unusual extended V-shaped dimeric structure with
CC       each monomer consisting of three distinct domains arranged along a
CC       curved 'spinal' alpha-helix. The N-terminal catalytic domain
CC       specifically recognizes the glutamate moiety of the substrate. The
CC       second domain is the NADPH-binding domain, and the third C-terminal
CC       domain is responsible for dimerization (By similarity). {ECO:0000250}.
CC   -!- MISCELLANEOUS: During catalysis, the active site Cys acts as a
CC       nucleophile attacking the alpha-carbonyl group of tRNA-bound glutamate
CC       with the formation of a thioester intermediate between enzyme and
CC       glutamate, and the concomitant release of tRNA(Glu). The thioester
CC       intermediate is finally reduced by direct hydride transfer from NADPH,
CC       to form the product GSA.
CC   -!- SIMILARITY: Belongs to the glutamyl-tRNA reductase family.
CC       {ECO:0000305}.
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DR   EMBL; M30785; AAA23953.1; -; Genomic_DNA.
DR   EMBL; M25323; AAA23954.1; -; Genomic_DNA.
DR   EMBL; X17434; CAA35476.1; -; Genomic_DNA.
DR   EMBL; U18555; AAC43436.1; -; Genomic_DNA.
DR   EMBL; U00096; AAC74294.1; -; Genomic_DNA.
DR   EMBL; AP009048; BAA36068.1; -; Genomic_DNA.
DR   EMBL; D10264; BAA20969.1; -; Genomic_DNA.
DR   PIR; A45918; BVECHA.
DR   RefSeq; NP_415728.1; NC_000913.3.
DR   RefSeq; WP_001299679.1; NZ_SSZK01000010.1.
DR   AlphaFoldDB; P0A6X1; -.
DR   SMR; P0A6X1; -.
DR   DIP; DIP-9877N; -.
DR   IntAct; P0A6X1; 1.
DR   STRING; 511145.b1210; -.
DR   ChEMBL; CHEMBL3309013; -.
DR   PaxDb; 511145-b1210; -.
DR   EnsemblBacteria; AAC74294; AAC74294; b1210.
DR   GeneID; 945777; -.
DR   KEGG; ecj:JW1201; -.
DR   KEGG; eco:b1210; -.
DR   PATRIC; fig|1411691.4.peg.1074; -.
DR   EchoBASE; EB0422; -.
DR   eggNOG; COG0373; Bacteria.
DR   HOGENOM; CLU_035113_2_2_6; -.
DR   InParanoid; P0A6X1; -.
DR   OMA; FAFKCAA; -.
DR   OrthoDB; 110209at2; -.
DR   PhylomeDB; P0A6X1; -.
DR   BioCyc; EcoCyc:GLUTRNAREDUCT-MONOMER; -.
DR   BioCyc; MetaCyc:GLUTRNAREDUCT-MONOMER; -.
DR   BRENDA; 1.2.1.70; 2026.
DR   SABIO-RK; P0A6X1; -.
DR   UniPathway; UPA00251; UER00316.
DR   PRO; PR:P0A6X1; -.
DR   Proteomes; UP000000318; Chromosome.
DR   Proteomes; UP000000625; Chromosome.
DR   GO; GO:0008883; F:glutamyl-tRNA reductase activity; IDA:EcoCyc.
DR   GO; GO:0042802; F:identical protein binding; IDA:EcoCyc.
DR   GO; GO:0050661; F:NADP binding; IEA:InterPro.
DR   GO; GO:0019353; P:protoporphyrinogen IX biosynthetic process from glutamate; IDA:EcoCyc.
DR   CDD; cd05213; NAD_bind_Glutamyl_tRNA_reduct; 1.
DR   Gene3D; 3.30.460.30; Glutamyl-tRNA reductase, N-terminal domain; 1.
DR   Gene3D; 3.40.50.720; NAD(P)-binding Rossmann-like Domain; 1.
DR   HAMAP; MF_00087; Glu_tRNA_reductase; 1.
DR   InterPro; IPR000343; 4pyrrol_synth_GluRdtase.
DR   InterPro; IPR015896; 4pyrrol_synth_GluRdtase_dimer.
DR   InterPro; IPR015895; 4pyrrol_synth_GluRdtase_N.
DR   InterPro; IPR018214; GluRdtase_CS.
DR   InterPro; IPR036453; GluRdtase_dimer_dom_sf.
DR   InterPro; IPR036343; GluRdtase_N_sf.
DR   InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR   InterPro; IPR006151; Shikm_DH/Glu-tRNA_Rdtase.
DR   NCBIfam; TIGR01035; hemA; 1.
DR   PANTHER; PTHR43013; GLUTAMYL-TRNA REDUCTASE; 1.
DR   PANTHER; PTHR43013:SF1; GLUTAMYL-TRNA REDUCTASE; 1.
DR   Pfam; PF00745; GlutR_dimer; 1.
DR   Pfam; PF05201; GlutR_N; 1.
DR   Pfam; PF01488; Shikimate_DH; 1.
DR   PIRSF; PIRSF000445; 4pyrrol_synth_GluRdtase; 1.
DR   SUPFAM; SSF69742; Glutamyl tRNA-reductase catalytic, N-terminal domain; 1.
DR   SUPFAM; SSF69075; Glutamyl tRNA-reductase dimerization domain; 1.
DR   SUPFAM; SSF51735; NAD(P)-binding Rossmann-fold domains; 1.
DR   PROSITE; PS00747; GLUTR; 1.
PE   1: Evidence at protein level;
KW   Direct protein sequencing; NADP; Oxidoreductase; Porphyrin biosynthesis;
KW   Reference proteome.
FT   CHAIN           1..418
FT                   /note="Glutamyl-tRNA reductase"
FT                   /id="PRO_0000114023"
FT   ACT_SITE        50
FT                   /note="Nucleophile"
FT   BINDING         49..52
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000305"
FT   BINDING         109
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000305"
FT   BINDING         114..116
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000305"
FT   BINDING         120
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000250"
FT   BINDING         189..194
FT                   /ligand="NADP(+)"
FT                   /ligand_id="ChEBI:CHEBI:58349"
FT                   /evidence="ECO:0000305"
FT   SITE            99
FT                   /note="Important for activity"
FT   MUTAGEN         7
FT                   /note="G->D: Loss of activity."
FT                   /evidence="ECO:0000269|PubMed:12370189"
FT   MUTAGEN         49
FT                   /note="T->V: 10% and 5% of wild-type reductase and esterase
FT                   activity, respectively."
FT                   /evidence="ECO:0000269|PubMed:17697121"
FT   MUTAGEN         50
FT                   /note="C->S: Loss of activity."
FT                   /evidence="ECO:0000269|PubMed:12370189"
FT   MUTAGEN         52
FT                   /note="R->K: 5% and 4% of wild-type reductase and esterase
FT                   activity, respectively."
FT                   /evidence="ECO:0000269|PubMed:17697121"
FT   MUTAGEN         52
FT                   /note="R->Q: Loss of activity."
FT                   /evidence="ECO:0000269|PubMed:17697121"
FT   MUTAGEN         54
FT                   /note="E->K: 6% and 2% of wild-type reductase and esterase
FT                   activity, respectively."
FT                   /evidence="ECO:0000269|PubMed:17697121"
FT   MUTAGEN         74
FT                   /note="C->S: No effect."
FT                   /evidence="ECO:0000269|PubMed:12370189"
FT   MUTAGEN         99
FT                   /note="H->N: 5% and 4% of wild-type reductase and esterase
FT                   activity, respectively."
FT                   /evidence="ECO:0000269|PubMed:17697121"
FT   MUTAGEN         106
FT                   /note="G->N: Loss of activity."
FT                   /evidence="ECO:0000269|PubMed:12370189"
FT   MUTAGEN         109
FT                   /note="S->A: 28% and 25% of wild-type reductase and
FT                   esterase activity, respectively."
FT                   /evidence="ECO:0000269|PubMed:17697121"
FT   MUTAGEN         114
FT                   /note="E->K: Loss of activity."
FT                   /evidence="ECO:0000269|PubMed:12370189"
FT   MUTAGEN         116
FT                   /note="Q->L: Loss of reductase activity. 30% of wild-type
FT                   esterase activity."
FT                   /evidence="ECO:0000269|PubMed:17697121"
FT   MUTAGEN         145
FT                   /note="S->F: Loss of activity."
FT                   /evidence="ECO:0000269|PubMed:12370189"
FT   MUTAGEN         170
FT                   /note="C->S: No effect."
FT                   /evidence="ECO:0000269|PubMed:12370189"
FT   MUTAGEN         191
FT                   /note="G->D: Loss of reductase activity. Retains esterase
FT                   activity."
FT                   /evidence="ECO:0000269|PubMed:12370189"
FT   MUTAGEN         314
FT                   /note="R->C: Loss of activity."
FT                   /evidence="ECO:0000269|PubMed:12370189"
FT   CONFLICT        151..168
FT                   /note="RVRTETDIGASAVSVAFA -> PFALKQISVPALCLSLLP (in Ref. 3;
FT                   CAA35476)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        172
FT                   /note="L -> V (in Ref. 3; CAA35476)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        240
FT                   /note="L -> M (in Ref. 3; CAA35476)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        243
FT                   /note="A -> R (in Ref. 1 and 4)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        365
FT                   /note="A -> R (in Ref. 3; CAA35476)"
FT                   /evidence="ECO:0000305"
SQ   SEQUENCE   418 AA;  46307 MW;  3CEE59AC53610A88 CRC64;
     MTLLALGINH KTAPVSLRER VSFSPDKLDQ ALDSLLAQPM VQGGVVLSTC NRTELYLSVE
     EQDNLQEALI RWLCDYHNLN EEDLRKSLYW HQDNDAVSHL MRVASGLDSL VLGEPQILGQ
     VKKAFADSQK GHMKASELER MFQKSFSVAK RVRTETDIGA SAVSVAFAAC TLARQIFESL
     STVTVLLVGA GETIELVARH LREHKVQKMI IANRTRERAQ ILADEVGAEV IALSDIDERL
     READIIISST ASPLPIIGKG MVERALKSRR NQPMLLVDIA VPRDVEPEVG KLANAYLYSV
     DDLQSIISHN LAQRKAAAVE AETIVAQETS EFMAWLRAQS ASETIREYRS QAEQVRDELT
     AKALAALEQG GDAQAIMQDL AWKLTNRLIH APTKSLQQAA RDGDNERLNI LRDSLGLE
//