Glutamyl-tRNA reductase - Escherichia coli (strain K12)

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P0A6X1 (HEM1_ECOLI) Reviewed, UniProtKB/Swiss-Prot

Last modified May 1, 2013. Version 80. History...

Clusters with 100%, 90%, 50% identity |

Documents (3) |

Third-party data

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Names·Attributes·General annotation·Ontologies·Sequence annotation·Sequences·References·Cross-refs·Entry info·Documents Customize order

Names and origin

Protein names

Recommended name:
Glutamyl-tRNA reductase
Short name=GluTR
EC=1.2.1.70

Gene names

Name:	hemA
Ordered Locus Names:	b1210, JW1201

Organism

Escherichia coli (strain K12) [Reference proteome] [HAMAP]

Taxonomic identifier

83333 [NCBI]

Taxonomic lineage

Bacteria › Proteobacteria › Gammaproteobacteria › Enterobacteriales › Enterobacteriaceae › Escherichia ›

Protein attributes

Sequence length	418 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Catalyzes the NADPH-dependent reduction of glutamyl-tRNA(Glu) to glutamate 1-semialdehyde (GSA). In the absence of NADPH, exhibits substrate esterase activity, leading to the release of glutamate from tRNA. Ref.9 Ref.10
Catalytic activity	L-glutamate 1-semialdehyde + NADP⁺ + tRNA(Glu) = L-glutamyl-tRNA(Glu) + NADPH. Ref.9
Enzyme regulation	Activated by Mg²⁺ ions. Inhibited by metal-chelating agents such as EDTA, EGTA, 1,10-phenanthroline, 2,2'-dipyridyl, and by PtCl4 and KPdCl4 as well as Ni²⁺ and Co²⁺. Also inhibited by iodoacetamide, N-tosyl-L-phenylalanine chloromethyl ketone, and 5,5'-dithiobis(2-nitrobenzoic acid), as well as glutamycin. Ref.9
Pathway	Porphyrin metabolism; protoporphyrin-IX biosynthesis; 5-aminolevulinate from L-glutamyl-tRNA(Glu): step 1/2. HAMAP-Rule MF_00087
Subunit structure	Homodimer. Interacts with glutamate-1-semialdehyde 2,1-aminomutase (GSA-AM), which forms a metabolic channeling between both enzymes to protect the reactive aldehyde species GSA. Ref.9 Ref.11
Domain	Possesses an unusual extended V-shaped dimeric structure with each monomer consisting of three distinct domains arranged along a curved 'spinal' alpha-helix. The N-terminal catalytic domain specifically recognizes the glutamate moiety of the substrate. The second domain is the NADPH-binding domain, and the third C-terminal domain is responsible for dimerization By similarity. HAMAP-Rule MF_00087
Miscellaneous	During catalysis, the active site Cys acts as a nucleophile attacking the alpha-carbonyl group of tRNA-bound glutamate with the formation of a thioester intermediate between enzyme and glutamate, and the concomitant release of tRNA(Glu). The thioester intermediate is finally reduced by direct hydride transfer from NADPH, to form the product GSA. HAMAP-Rule MF_00087
Sequence similarities	Belongs to the glutamyl-tRNA reductase family.
Biophysicochemical properties	Kinetic parameters: K_M=24 µM for L-glutamyl-tRNA(Glu) Ref.9 K_M=39 µM for NADPH

Ontologies

Keywords
Biological process	Porphyrin biosynthesis
Ligand	NADP
Molecular function	Oxidoreductase
Technical term	Complete proteome Direct protein sequencing Reference proteome
Gene Ontology (GO)
Biological_process	chlorophyll biosynthetic process Inferred from electronic annotation. Source: HAMAP protoporphyrinogen IX biosynthetic process Inferred from electronic annotation. Source: UniProtKB-UniPathway
Molecular_function	NADP binding Inferred from electronic annotation. Source: InterPro glutamyl-tRNA reductase activity Inferred from electronic annotation. Source: HAMAP
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 418

418

Glutamyl-tRNA reductase HAMAP-Rule MF_00087

PRO_0000114023

Regions

Nucleotide binding

189 – 194

NADP Probable

Region

49 – 52

Substrate binding Probable

Region

114 – 116

Substrate binding Probable

Sites

Active site

Nucleophile

Binding site

109

Substrate Probable

Binding site

120

Substrate By similarity

Site

Important for activity

Experimental info

Mutagenesis

G → D: Loss of activity. Ref.9

Mutagenesis

T → V: 10% and 5% of wild-type reductase and esterase activity, respectively. Ref.12

Mutagenesis

C → S: Loss of activity. Ref.9

Mutagenesis

R → K: 5% and 4% of wild-type reductase and esterase activity, respectively. Ref.12

Mutagenesis

R → Q: Loss of activity. Ref.12

Mutagenesis

E → K: 6% and 2% of wild-type reductase and esterase activity, respectively. Ref.12

Mutagenesis

C → S: No effect. Ref.9

Mutagenesis

H → N: 5% and 4% of wild-type reductase and esterase activity, respectively. Ref.12

Mutagenesis

106

G → N: Loss of activity. Ref.9

Mutagenesis

109

S → A: 28% and 25% of wild-type reductase and esterase activity, respectively. Ref.12

Mutagenesis

114

E → K: Loss of activity. Ref.9

Mutagenesis

116

Q → L: Loss of reductase activity. 30% of wild-type esterase activity. Ref.12

Mutagenesis

145

S → F: Loss of activity. Ref.9

Mutagenesis

170

C → S: No effect. Ref.9

Mutagenesis

191

G → D: Loss of reductase activity. Retains esterase activity. Ref.9

Mutagenesis

314

R → C: Loss of activity. Ref.9

Sequence conflict

151 – 168

RVRTE…SVAFA → PFALKQISVPALCLSLLP in CAA35476. Ref.3

Sequence conflict

172

L → V in CAA35476. Ref.3

Sequence conflict

240

L → M in CAA35476. Ref.3

Sequence conflict

243

A → R Ref.1

Sequence conflict

243

A → R Ref.4

Sequence conflict

365

A → R in CAA35476. Ref.3

Sequences

Sequence

Length

Mass (Da)

Tools

P0A6X1 [UniParc].

Last modified March 29, 2005. Version 1.
Checksum: 3CEE59AC53610A88
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FASTA

418

46,307

        10         20         30         40         50         60 
MTLLALGINH KTAPVSLRER VSFSPDKLDQ ALDSLLAQPM VQGGVVLSTC NRTELYLSVE 

        70         80         90        100        110        120 
EQDNLQEALI RWLCDYHNLN EEDLRKSLYW HQDNDAVSHL MRVASGLDSL VLGEPQILGQ 

       130        140        150        160        170        180 
VKKAFADSQK GHMKASELER MFQKSFSVAK RVRTETDIGA SAVSVAFAAC TLARQIFESL 

       190        200        210        220        230        240 
STVTVLLVGA GETIELVARH LREHKVQKMI IANRTRERAQ ILADEVGAEV IALSDIDERL 

       250        260        270        280        290        300 
READIIISST ASPLPIIGKG MVERALKSRR NQPMLLVDIA VPRDVEPEVG KLANAYLYSV 

       310        320        330        340        350        360 
DDLQSIISHN LAQRKAAAVE AETIVAQETS EFMAWLRAQS ASETIREYRS QAEQVRDELT 

       370        380        390        400        410 
AKALAALEQG GDAQAIMQDL AWKLTNRLIH APTKSLQQAA RDGDNERLNI LRDSLGLE

« Hide

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Cloning and structure of the hem A gene of Escherichia coli K-12." Li J.-M., Russell C.S., Cosloy S.D. Gene 82:209-217(1989) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA]. Strain: K12.
[2]	"Isolation, nucleotide sequence, and preliminary characterization of the Escherichia coli K-12 hemA gene." Verkamp E., Chelm B.K. J. Bacteriol. 171:4728-4735(1989) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA]. Strain: K12.
[3]	"Isolation and nucleotide sequence of the hemA gene of Escherichia coli K12." Drolet M., Peloquin L., Echelard Y., Cousineau L., Sasarman A. Mol. Gen. Genet. 216:347-352(1989) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA]. Strain: K12.
[4]	"Expression of genes kdsA and kdsB involved in 3-deoxy-D-manno-octulosonic acid metabolism and biosynthesis of enterobacterial lipopolysaccharide is growth phase regulated primarily at the transcriptional level in Escherichia coli K-12." Strohmaier H., Remler P., Renner W., Hoegenauer G. J. Bacteriol. 177:4488-4500(1995) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA]. Strain: K12.
[5]	"A 718-kb DNA sequence of the Escherichia coli K-12 genome corresponding to the 12.7-28.0 min region on the linkage map." Oshima T., Aiba H., Baba T., Fujita K., Hayashi K., Honjo A., Ikemoto K., Inada T., Itoh T., Kajihara M., Kanai K., Kashimoto K., Kimura S., Kitagawa M., Makino K., Masuda S., Miki T., Mizobuchi K. Horiuchi T. DNA Res. 3:137-155(1996) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[6]	"The complete genome sequence of Escherichia coli K-12." Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V., Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F., Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B., Shao Y. Science 277:1453-1474(1997) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / MG1655 / ATCC 47076.
[7]	"Highly accurate genome sequences of Escherichia coli K-12 strains MG1655 and W3110." Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S., Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T. Mol. Syst. Biol. 2:E1-E5(2006) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[8]	"Cloning and characterization of genes involved in the biosynthesis of delta-aminolevulinic acid in Escherichia coli." Ikemi M., Murakami K., Hashimoto M., Murooka Y. Gene 121:127-132(1992) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-49.
[9]	"Escherichia coli glutamyl-tRNA reductase. Trapping the thioester intermediate." Schauer S., Chaturvedi S., Randau L., Moser J., Kitabatake M., Lorenz S., Verkamp E., Schubert W.-D., Nakayashiki T., Murai M., Wall K., Thomann H.-U., Heinz D.W., Inokuchi H., Soell D., Jahn D. J. Biol. Chem. 277:48657-48663(2002) [PubMed] [Europe PMC] [Abstract] Cited for: PROTEIN SEQUENCE OF 1-15, FUNCTION, CATALYTIC ACTIVITY, CHARACTERIZATION, ENZYME REGULATION, MASS SPECTROMETRY, SUBUNIT, KINETIC PARAMETERS, CATALYTIC MECHANISM, MUTAGENESIS OF GLY-7; CYS-50; CYS-74; GLY-106; GLU-114; SER-145; CYS-170; GLY-191 AND ARG-314.
[10]	"Glutamyl-tRNA reductase from Escherichia coli and Synechocystis 6803. Gene structure and expression." Verkamp E., Jahn M., Jahn D., Kumar A.M., Soell D. J. Biol. Chem. 267:8275-8280(1992) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION.
[11]	"Complex formation between glutamyl-tRNA reductase and glutamate-1-semialdehyde 2,1-aminomutase in Escherichia coli during the initial reactions of porphyrin biosynthesis." Lueer C., Schauer S., Moebius K., Schulze J., Schubert W.-D., Heinz D.W., Jahn D., Moser J. J. Biol. Chem. 280:18568-18572(2005) [PubMed] [Europe PMC] [Abstract] Cited for: INTERACTION WITH GSA-AM.
[12]	"Glutamate recognition and hydride transfer by Escherichia coli glutamyl-tRNA reductase." Lueer C., Schauer S., Virus S., Schubert W.-D., Heinz D.W., Moser J., Jahn D. FEBS J. 274:4609-4614(2007) [PubMed] [Europe PMC] [Abstract] Cited for: MUTAGENESIS OF THR-49; ARG-52; GLU-54; HIS-99; SER-109 AND GLN-116.

Cross-references

Sequence databases
EMBL GenBank DDBJ	M30785 Genomic DNA. Translation: AAA23953.1. M25323 Genomic DNA. Translation: AAA23954.1. X17434 Genomic DNA. Translation: CAA35476.1. U18555 Genomic DNA. Translation: AAC43436.1. U00096 Genomic DNA. Translation: AAC74294.1. AP009048 Genomic DNA. Translation: BAA36068.1. D10264 Genomic DNA. Translation: BAA20969.1.
PIR	BVECHA. A45918.
RefSeq	NP_415728.1. NC_000913.2. YP_489477.1. NC_007779.1.
3D structure databases
ProteinModelPortal	P0A6X1.
SMR	P0A6X1. Positions 3-398.
ModBase	Search...
Protein-protein interaction databases
IntAct	P0A6X1. 1 interaction.
STRING	511145.b1210.
Proteomic databases
PaxDb	P0A6X1.
PRIDE	P0A6X1.
Protocols and materials databases
StructuralBiologyKnowledgebase	Search...
Genome annotation databases
EnsemblBacteria	AAC74294; AAC74294; b1210. BAA36068; BAA36068; BAA36068.
GeneID	12931822. 945777.
KEGG	ecj:Y75_p1182. eco:b1210.
PATRIC	32117672. VBIEscCol129921_1258.
Organism-specific databases
EchoBASE	EB0422.
EcoGene	EG10427. hemA.
Phylogenomic databases
eggNOG	COG0373.
HOGENOM	HOG000109650.
KO	K02492.
OMA	GPILNRL.
ProtClustDB	PRK00045.
Enzyme and pathway databases
BioCyc	EcoCyc:GLUTRNAREDUCT-MONOMER. ECOL316407:JW1201-MONOMER. MetaCyc:GLUTRNAREDUCT-MONOMER.
UniPathway	UPA00251; UER00316.
Gene expression databases
Genevestigator	P0A6X1.
Family and domain databases
Gene3D	3.40.50.720. 1 hit.
HAMAP	MF_00087. Glu-tRNA_reductase.
InterPro	IPR000343. 4pyrrol_synth_GluRdtase. IPR015896. 4pyrrol_synth_GluRdtase_dimer. IPR015895. 4pyrrol_synth_GluRdtase_N. IPR016040. NAD(P)-bd_dom. IPR018214. Pyrrol_synth_GluRdtase_CS. IPR006151. Shikm_DH/Glu-tRNA_Rdtase. [Graphical view]
Pfam	PF00745. GlutR_dimer. 1 hit. PF05201. GlutR_N. 1 hit. PF01488. Shikimate_DH. 1 hit. [Graphical view]
PIRSF	PIRSF000445. 4pyrrol_synth_GluRdtase. 1 hit.
SUPFAM	SSF69075. 4pyrrol_synth_GluRdtase_C. 1 hit. SSF69742. GlutR. 1 hit.
TIGRFAMs	TIGR01035. hemA. 1 hit.
PROSITE	PS00747. GLUTR. 1 hit. [Graphical view]
ProtoNet	Search...

Entry information

Entry name

HEM1_ECOLI

Accession

Primary (citable) accession number: P0A6X1
Secondary accession number(s): P13580, Q59405

Entry history

Integrated into UniProtKB/Swiss-Prot:	March 29, 2005
Last sequence update:	March 29, 2005
Last modified:	May 1, 2013
This is version 80 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program

Relevant documents

Escherichia coli

Escherichia coli (strain K12): entries and cross-references to EcoGene

PATHWAY comments

Index of metabolic and biosynthesis pathways

SIMILARITY comments

Index of protein domains and families

Names·Attributes·General annotation·Ontologies·Sequence annotation·Sequences·References·Cross-refs·Entry info·Documents

Preferred order of sections

Names and origin

Protein attributes

General annotation (Comments)

Ontologies

Sequence annotation (Features)

Molecule processing

Regions

Sites

Experimental info

Sequences

References

Cross-references

Sequence databases

3D structure databases

Protein-protein interaction databases

Proteomic databases

Protocols and materials databases

Genome annotation databases

Organism-specific databases

Phylogenomic databases

Enzyme and pathway databases

Gene expression databases

Family and domain databases

Entry information

Relevant documents