ID   GSHB_ECOLI              Reviewed;         316 AA.
AC   P04425; Q2M9P6;
DT   13-AUG-1987, integrated into UniProtKB/Swiss-Prot.
DT   13-AUG-1987, sequence version 1.
DT   27-MAR-2024, entry version 198.
DE   RecName: Full=Glutathione synthetase;
DE            EC=6.3.2.3;
DE   AltName: Full=GSH synthetase;
DE            Short=GSH-S;
DE            Short=GSHase;
DE   AltName: Full=Glutathione synthase;
GN   Name=gshB; Synonyms=gsh-II; OrderedLocusNames=b2947, JW2914;
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=B;
RX   PubMed=6393055; DOI=10.1093/nar/12.24.9299;
RA   Gushima H., Yasuda S., Soeda E., Yokota M., Kondo M., Kimura A.;
RT   "Complete nucleotide sequence of the E. coli glutathione synthetase gsh-
RT   II.";
RL   Nucleic Acids Res. 12:9299-9307(1984).
RN   [2]
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   [3]
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   [4]
RP   X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS).
RC   STRAIN=B;
RX   PubMed=8445637; DOI=10.1006/jmbi.1993.1106;
RA   Yamaguchi H., Kato H., Hata Y., Nishioka T., Kimura A., Oda J., Katsube Y.;
RT   "Three-dimensional structure of the glutathione synthetase from Escherichia
RT   coli B at 2.0-A resolution.";
RL   J. Mol. Biol. 229:1083-1100(1993).
RN   [5]
RP   X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS).
RC   STRAIN=B;
RX   PubMed=9010922; DOI=10.1093/protein/9.12.1083;
RA   Matsuda K., Mizuguchi K., Nishioka T., Kato H., Go N., Oda J.;
RT   "Crystal structure of glutathione synthetase at optimal pH: domain
RT   architecture and structural similarity with other proteins.";
RL   Protein Eng. 9:1083-1092(1996).
RN   [6]
RP   MUTAGENESIS OF PRO-227 AND GLY-240.
RX   PubMed=8241129; DOI=10.1021/bi00097a018;
RA   Tanaka T., Yamaguchi H., Kato H., Nishioka T., Katsube Y., Oda J.;
RT   "Flexibility impaired by mutations revealed the multifunctional roles of
RT   the loop in glutathione synthetase.";
RL   Biochemistry 32:12398-12404(1993).
RN   [7]
RP   PROTEIN SEQUENCE OF 234-242, AND MUTAGENESIS OF ARG-233 AND ARG-241.
RC   STRAIN=B;
RX   PubMed=1540581; DOI=10.1021/bi00123a007;
RA   Tanaka T., Kato H., Nishioka T., Oda J.;
RT   "Mutational and proteolytic studies on a flexible loop in glutathione
RT   synthetase from Escherichia coli B: the loop and arginine 233 are critical
RT   for the catalytic reaction.";
RL   Biochemistry 31:2259-2265(1992).
RN   [8]
RP   MUTAGENESIS OF CYSTEINE RESIDUES.
RC   STRAIN=B;
RX   PubMed=3042775; DOI=10.1016/s0021-9258(18)37833-5;
RA   Kato H., Tanaka T., Nishioka T., Kimura A., Oda J.;
RT   "Role of cysteine residues in glutathione synthetase from Escherichia coli
RT   B. Chemical modification and oligonucleotide site-directed mutagenesis.";
RL   J. Biol. Chem. 263:11646-11651(1988).
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=ATP + gamma-L-glutamyl-L-cysteine + glycine = ADP +
CC         glutathione + H(+) + phosphate; Xref=Rhea:RHEA:13557,
CC         ChEBI:CHEBI:15378, ChEBI:CHEBI:30616, ChEBI:CHEBI:43474,
CC         ChEBI:CHEBI:57305, ChEBI:CHEBI:57925, ChEBI:CHEBI:58173,
CC         ChEBI:CHEBI:456216; EC=6.3.2.3;
CC   -!- COFACTOR:
CC       Name=Mg(2+); Xref=ChEBI:CHEBI:18420; Evidence={ECO:0000250};
CC       Name=Mn(2+); Xref=ChEBI:CHEBI:29035; Evidence={ECO:0000250};
CC       Note=Binds 1 Mg(2+) or Mn(2+) ion per subunit. {ECO:0000250};
CC   -!- ACTIVITY REGULATION: Inhibited by 7,8-dihydrofolate, methotrexate and
CC       trimethoprim.
CC   -!- PATHWAY: Sulfur metabolism; glutathione biosynthesis; glutathione from
CC       L-cysteine and L-glutamate: step 2/2.
CC   -!- SUBUNIT: Homotetramer.
CC   -!- SIMILARITY: Belongs to the prokaryotic GSH synthase family.
CC       {ECO:0000305}.
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DR   EMBL; X01666; CAA25826.1; -; Genomic_DNA.
DR   EMBL; U28377; AAA69114.1; -; Genomic_DNA.
DR   EMBL; U00096; AAC75984.1; -; Genomic_DNA.
DR   EMBL; AP009048; BAE77010.1; -; Genomic_DNA.
DR   PIR; A01194; SYECGS.
DR   RefSeq; NP_417422.1; NC_000913.3.
DR   RefSeq; WP_000593273.1; NZ_STEB01000001.1.
DR   PDB; 1GLV; X-ray; 2.70 A; A=1-316.
DR   PDB; 1GSA; X-ray; 2.00 A; A=1-316.
DR   PDB; 1GSH; X-ray; 2.00 A; A=1-316.
DR   PDB; 2GLT; X-ray; 2.20 A; A=1-316.
DR   PDBsum; 1GLV; -.
DR   PDBsum; 1GSA; -.
DR   PDBsum; 1GSH; -.
DR   PDBsum; 2GLT; -.
DR   AlphaFoldDB; P04425; -.
DR   SMR; P04425; -.
DR   BioGRID; 4259406; 20.
DR   BioGRID; 851765; 1.
DR   DIP; DIP-9840N; -.
DR   IntAct; P04425; 5.
DR   STRING; 511145.b2947; -.
DR   jPOST; P04425; -.
DR   PaxDb; 511145-b2947; -.
DR   EnsemblBacteria; AAC75984; AAC75984; b2947.
DR   GeneID; 75205218; -.
DR   GeneID; 947445; -.
DR   KEGG; ecj:JW2914; -.
DR   KEGG; eco:b2947; -.
DR   PATRIC; fig|1411691.4.peg.3786; -.
DR   EchoBASE; EB0414; -.
DR   eggNOG; COG0189; Bacteria.
DR   HOGENOM; CLU_068239_0_0_6; -.
DR   InParanoid; P04425; -.
DR   OMA; IWMRKDP; -.
DR   OrthoDB; 9785415at2; -.
DR   PhylomeDB; P04425; -.
DR   BioCyc; EcoCyc:GLUTATHIONE-SYN-MONOMER; -.
DR   BioCyc; MetaCyc:GLUTATHIONE-SYN-MONOMER; -.
DR   BRENDA; 6.3.2.3; 2026.
DR   UniPathway; UPA00142; UER00210.
DR   EvolutionaryTrace; P04425; -.
DR   PRO; PR:P04425; -.
DR   Proteomes; UP000000318; Chromosome.
DR   Proteomes; UP000000625; Chromosome.
DR   GO; GO:0005737; C:cytoplasm; IBA:GO_Central.
DR   GO; GO:0005829; C:cytosol; IDA:EcoCyc.
DR   GO; GO:0005524; F:ATP binding; IEA:UniProtKB-UniRule.
DR   GO; GO:0004363; F:glutathione synthase activity; IDA:EcoCyc.
DR   GO; GO:0042802; F:identical protein binding; IDA:EcoCyc.
DR   GO; GO:0000287; F:magnesium ion binding; IDA:EcoCyc.
DR   GO; GO:0006750; P:glutathione biosynthetic process; IMP:EcoCyc.
DR   GO; GO:0051289; P:protein homotetramerization; IDA:EcoCyc.
DR   Gene3D; 3.40.50.20; -; 1.
DR   Gene3D; 3.30.1490.20; ATP-grasp fold, A domain; 1.
DR   Gene3D; 3.30.470.20; ATP-grasp fold, B domain; 1.
DR   HAMAP; MF_00162; GSH_S; 1.
DR   InterPro; IPR011761; ATP-grasp.
DR   InterPro; IPR013815; ATP_grasp_subdomain_1.
DR   InterPro; IPR006284; Glut_synth_pro.
DR   InterPro; IPR004218; GSHS_ATP-bd.
DR   InterPro; IPR004215; GSHS_N.
DR   InterPro; IPR016185; PreATP-grasp_dom_sf.
DR   NCBIfam; TIGR01380; glut_syn; 1.
DR   PANTHER; PTHR21621:SF4; GLUTATHIONE SYNTHETASE; 1.
DR   PANTHER; PTHR21621; RIBOSOMAL PROTEIN S6 MODIFICATION PROTEIN; 1.
DR   Pfam; PF02955; GSH-S_ATP; 1.
DR   Pfam; PF02951; GSH-S_N; 1.
DR   SUPFAM; SSF56059; Glutathione synthetase ATP-binding domain-like; 1.
DR   SUPFAM; SSF52440; PreATP-grasp domain; 1.
DR   PROSITE; PS50975; ATP_GRASP; 1.
PE   1: Evidence at protein level;
KW   3D-structure; ATP-binding; Direct protein sequencing;
KW   Glutathione biosynthesis; Ligase; Magnesium; Manganese; Metal-binding;
KW   Nucleotide-binding; Reference proteome.
FT   CHAIN           1..316
FT                   /note="Glutathione synthetase"
FT                   /id="PRO_0000197465"
FT   DOMAIN          125..310
FT                   /note="ATP-grasp"
FT   BINDING         151..207
FT                   /ligand="ATP"
FT                   /ligand_id="ChEBI:CHEBI:30616"
FT                   /evidence="ECO:0000250"
FT   BINDING         281
FT                   /ligand="Mg(2+)"
FT                   /ligand_id="ChEBI:CHEBI:18420"
FT                   /evidence="ECO:0000250"
FT   BINDING         283
FT                   /ligand="Mg(2+)"
FT                   /ligand_id="ChEBI:CHEBI:18420"
FT                   /evidence="ECO:0000250"
FT   MUTAGEN         122
FT                   /note="C->A: No loss of activity."
FT                   /evidence="ECO:0000269|PubMed:3042775"
FT   MUTAGEN         195
FT                   /note="C->A: No loss of activity."
FT                   /evidence="ECO:0000269|PubMed:3042775"
FT   MUTAGEN         222
FT                   /note="C->A: No loss of activity."
FT                   /evidence="ECO:0000269|PubMed:3042775"
FT   MUTAGEN         227
FT                   /note="P->V: Loss of activity."
FT                   /evidence="ECO:0000269|PubMed:8241129"
FT   MUTAGEN         233
FT                   /note="R->A,K: Loss of activity."
FT                   /evidence="ECO:0000269|PubMed:1540581"
FT   MUTAGEN         240
FT                   /note="G->V: Loss of activity."
FT                   /evidence="ECO:0000269|PubMed:8241129"
FT   MUTAGEN         241
FT                   /note="R->A,K: No loss of activity."
FT                   /evidence="ECO:0000269|PubMed:1540581"
FT   MUTAGEN         289
FT                   /note="C->A: No loss of activity."
FT                   /evidence="ECO:0000269|PubMed:3042775"
FT   STRAND          3..7
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   HELIX           11..13
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   TURN            16..18
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   HELIX           20..30
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   STRAND          34..38
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   HELIX           40..42
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   STRAND          43..46
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   STRAND          49..59
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   STRAND          67..76
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   HELIX           77..79
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   STRAND          80..85
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   HELIX           93..107
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   STRAND          111..114
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   HELIX           116..121
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   HELIX           126..131
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   TURN            132..134
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   STRAND          138..142
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   HELIX           144..154
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   STRAND          155..160
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   TURN            166..169
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   STRAND          171..173
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   HELIX           180..187
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   TURN            188..192
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   STRAND          195..199
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   HELIX           202..206
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   STRAND          208..214
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   STRAND          220..226
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   HELIX           236..238
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   STRAND          241..246
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   HELIX           249..264
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   STRAND          269..275
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   STRAND          278..283
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   HELIX           290..296
FT                   /evidence="ECO:0007829|PDB:1GSA"
FT   HELIX           301..312
FT                   /evidence="ECO:0007829|PDB:1GSA"
SQ   SEQUENCE   316 AA;  35561 MW;  F2E67AB29DE113DD CRC64;
     MIKLGIVMDP IANINIKKDS SFAMLLEAQR RGYELHYMEM GDLYLINGEA RAHTRTLNVK
     QNYEEWFSFV GEQDLPLADL DVILMRKDPP FDTEFIYATY ILERAEEKGT LIVNKPQSLR
     DCNEKLFTAW FSDLTPETLV TRNKAQLKAF WEKHSDIILK PLDGMGGASI FRVKEGDPNL
     GVIAETLTEH GTRYCMAQNY LPAIKDGDKR VLVVDGEPVP YCLARIPQGG ETRGNLAAGG
     RGEPRPLTES DWKIARQIGP TLKEKGLIFV GLDIIGDRLT EINVTSPTCI REIEAEFPVS
     ITGMLMDAIE ARLQQQ
//