HEM1_ECOLI
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 03-AUG-2022, entry version 135.
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; Proteobacteria; 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; P0A6X1; -.
DR PRIDE; P0A6X1; -.
DR EnsemblBacteria; AAC74294; AAC74294; b1210.
DR EnsemblBacteria; BAA36068; BAA36068; BAA36068.
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 PhylomeDB; P0A6X1; -.
DR BioCyc; EcoCyc:GLUTRNAREDUCT-MON; -.
DR BioCyc; MetaCyc:GLUTRNAREDUCT-MON; -.
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 Gene3D; 3.30.460.30; -; 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 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; SSF51735; SSF51735; 1.
DR SUPFAM; SSF69075; SSF69075; 1.
DR SUPFAM; SSF69742; SSF69742; 1.
DR TIGRFAMs; TIGR01035; hemA; 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
