GSHB_ECOLI
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 03-AUG-2022, entry version 190.
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; Proteobacteria; 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; P04425; -.
DR PRIDE; P04425; -.
DR EnsemblBacteria; AAC75984; AAC75984; b2947.
DR EnsemblBacteria; BAE77010; BAE77010; BAE77010.
DR GeneID; 66673172; -.
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; WMRKDPP; -.
DR PhylomeDB; P04425; -.
DR BioCyc; EcoCyc:GLUTATHIONE-SYN-MON; -.
DR BioCyc; MetaCyc:GLUTATHIONE-SYN-MON; -.
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:0016879; F:ligase activity, forming carbon-nitrogen bonds; IBA:GO_Central.
DR GO; GO:0000287; F:magnesium ion binding; IDA:EcoCyc.
DR GO; GO:0006750; P:glutathione biosynthetic process; IMP:EcoCyc.
DR Gene3D; 3.30.1490.20; -; 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 PANTHER; PTHR21621:SF4; PTHR21621:SF4; 1.
DR Pfam; PF02955; GSH-S_ATP; 1.
DR Pfam; PF02951; GSH-S_N; 1.
DR SUPFAM; SSF52440; SSF52440; 1.
DR TIGRFAMs; TIGR01380; glut_syn; 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
