ID   GSP_SHIFL               Reviewed;         619 AA.
AC   P0AES1; P43675;
DT   20-DEC-2005, integrated into UniProtKB/Swiss-Prot.
DT   20-DEC-2005, sequence version 1.
DT   27-MAR-2024, entry version 114.
DE   RecName: Full=Bifunctional glutathionylspermidine synthetase/amidase;
DE            Short=GspSA;
DE   Includes:
DE     RecName: Full=Glutathionylspermidine amidase;
DE              Short=Gsp amidase;
DE              EC=3.5.1.78 {ECO:0000250|UniProtKB:P0AES0};
DE     AltName: Full=Glutathionylspermidine amidohydrolase [spermidine-forming];
DE   Includes:
DE     RecName: Full=Glutathionylspermidine synthetase;
DE              Short=Gsp synthetase;
DE              EC=6.3.1.8 {ECO:0000250|UniProtKB:P0AES0};
DE     AltName: Full=Glutathione:spermidine ligase [ADP-forming];
DE     AltName: Full=Gsp synthase;
GN   Name=gss; Synonyms=gsp; OrderedLocusNames=SF3035, S3236;
OS   Shigella flexneri.
OC   Bacteria; Pseudomonadota; Gammaproteobacteria; Enterobacterales;
OC   Enterobacteriaceae; Shigella.
OX   NCBI_TaxID=623;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=301 / Serotype 2a;
RX   PubMed=12384590; DOI=10.1093/nar/gkf566;
RA   Jin Q., Yuan Z., Xu J., Wang Y., Shen Y., Lu W., Wang J., Liu H., Yang J.,
RA   Yang F., Zhang X., Zhang J., Yang G., Wu H., Qu D., Dong J., Sun L.,
RA   Xue Y., Zhao A., Gao Y., Zhu J., Kan B., Ding K., Chen S., Cheng H.,
RA   Yao Z., He B., Chen R., Ma D., Qiang B., Wen Y., Hou Y., Yu J.;
RT   "Genome sequence of Shigella flexneri 2a: insights into pathogenicity
RT   through comparison with genomes of Escherichia coli K12 and O157.";
RL   Nucleic Acids Res. 30:4432-4441(2002).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=ATCC 700930 / 2457T / Serotype 2a;
RX   PubMed=12704152; DOI=10.1128/iai.71.5.2775-2786.2003;
RA   Wei J., Goldberg M.B., Burland V., Venkatesan M.M., Deng W., Fournier G.,
RA   Mayhew G.F., Plunkett G. III, Rose D.J., Darling A., Mau B., Perna N.T.,
RA   Payne S.M., Runyen-Janecky L.J., Zhou S., Schwartz D.C., Blattner F.R.;
RT   "Complete genome sequence and comparative genomics of Shigella flexneri
RT   serotype 2a strain 2457T.";
RL   Infect. Immun. 71:2775-2786(2003).
CC   -!- FUNCTION: Catalyzes the formation of an amide bond between glutathione
CC       (GSH) and spermidine coupled with hydrolysis of ATP; also catalyzes the
CC       opposing reaction, i.e. the hydrolysis of glutathionylspermidine (Gsp)
CC       back to glutathione and spermidine. May act synergistically with
CC       glutaredoxin to regulate the redox environment and defend against
CC       oxidative damage. {ECO:0000250|UniProtKB:P0AES0}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=ATP + glutathione + spermidine = ADP + glutathionylspermidine
CC         + H(+) + phosphate; Xref=Rhea:RHEA:21272, ChEBI:CHEBI:15378,
CC         ChEBI:CHEBI:30616, ChEBI:CHEBI:43474, ChEBI:CHEBI:57834,
CC         ChEBI:CHEBI:57835, ChEBI:CHEBI:57925, ChEBI:CHEBI:456216; EC=6.3.1.8;
CC         Evidence={ECO:0000250|UniProtKB:P0AES0};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=glutathionylspermidine + H2O = glutathione + spermidine;
CC         Xref=Rhea:RHEA:17173, ChEBI:CHEBI:15377, ChEBI:CHEBI:57834,
CC         ChEBI:CHEBI:57835, ChEBI:CHEBI:57925; EC=3.5.1.78;
CC         Evidence={ECO:0000250|UniProtKB:P0AES0};
CC   -!- ACTIVITY REGULATION: When exposed to oxidative stress, Gsp amidase
CC       activity is transiently inhibited in vivo by oxidation of the catalytic
CC       Cys-59 thiol to sulfenic acid; this modification does not affect Gsp
CC       synthetase activity. {ECO:0000250|UniProtKB:P0AES0}.
CC   -!- PATHWAY: Sulfur metabolism; glutathione metabolism.
CC   -!- PATHWAY: Amine and polyamine metabolism; spermidine metabolism.
CC   -!- SUBUNIT: Homodimer. {ECO:0000250|UniProtKB:P0AES0}.
CC   -!- DOMAIN: The two activities reside in distinct domains (N-terminal
CC       amidase and C-terminal synthetase). The two domains expressed
CC       independently are folded and functional (By similarity). {ECO:0000250}.
CC   -!- PTM: Oxidation of Cys-59 to sulfenic acid during oxidative stress
CC       selectively inhibits the amidase activity.
CC       {ECO:0000250|UniProtKB:P0AES0}.
CC   -!- SIMILARITY: In the C-terminal section; belongs to the
CC       glutathionylspermidine synthase preATP-grasp family. {ECO:0000305}.
CC   ---------------------------------------------------------------------------
CC   Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC   Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC   ---------------------------------------------------------------------------
DR   EMBL; AE005674; AAN44513.2; -; Genomic_DNA.
DR   EMBL; AE014073; AAP18324.1; -; Genomic_DNA.
DR   RefSeq; NP_708806.2; NC_004337.2.
DR   RefSeq; WP_001297309.1; NZ_WPGW01000034.1.
DR   AlphaFoldDB; P0AES1; -.
DR   SMR; P0AES1; -.
DR   STRING; 198214.SF3035; -.
DR   PaxDb; 198214-SF3035; -.
DR   GeneID; 1026646; -.
DR   GeneID; 75205173; -.
DR   KEGG; sfl:SF3035; -.
DR   KEGG; sfx:S3236; -.
DR   PATRIC; fig|198214.7.peg.3609; -.
DR   HOGENOM; CLU_478805_0_0_6; -.
DR   UniPathway; UPA00204; -.
DR   UniPathway; UPA00819; -.
DR   Proteomes; UP000001006; Chromosome.
DR   Proteomes; UP000002673; Chromosome.
DR   GO; GO:0005524; F:ATP binding; IEA:UniProtKB-KW.
DR   GO; GO:0008884; F:glutathionylspermidine amidase activity; IEA:UniProtKB-EC.
DR   GO; GO:0008885; F:glutathionylspermidine synthase activity; IEA:UniProtKB-EC.
DR   GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR   GO; GO:0006749; P:glutathione metabolic process; IEA:UniProtKB-UniPathway.
DR   GO; GO:0008216; P:spermidine metabolic process; IEA:UniProtKB-UniPathway.
DR   Gene3D; 3.30.1490.330; -; 1.
DR   Gene3D; 3.90.1720.10; endopeptidase domain like (from Nostoc punctiforme); 1.
DR   InterPro; IPR007921; CHAP_dom.
DR   InterPro; IPR005494; GSPS_pre-ATP-grasp-like_dom.
DR   InterPro; IPR038765; Papain-like_cys_pep_sf.
DR   InterPro; IPR016185; PreATP-grasp_dom_sf.
DR   PANTHER; PTHR30094; BIFUNCTIONAL GLUTATHIONYLSPERMIDINE SYNTHETASE/AMIDASE-RELATED; 1.
DR   PANTHER; PTHR30094:SF0; BIFUNCTIONAL GLUTATHIONYLSPERMIDINE SYNTHETASE_AMIDASE-RELATED; 1.
DR   Pfam; PF05257; CHAP; 1.
DR   Pfam; PF03738; GSP_synth; 1.
DR   SUPFAM; SSF54001; Cysteine proteinases; 1.
DR   SUPFAM; SSF56059; Glutathione synthetase ATP-binding domain-like; 1.
DR   SUPFAM; SSF52440; PreATP-grasp domain; 1.
DR   PROSITE; PS50911; CHAP; 1.
PE   3: Inferred from homology;
KW   ATP-binding; Hydrolase; Ligase; Magnesium; Metal-binding;
KW   Multifunctional enzyme; Nucleotide-binding; Oxidation; Reference proteome.
FT   INIT_MET        1
FT                   /note="Removed"
FT                   /evidence="ECO:0000250"
FT   CHAIN           2..619
FT                   /note="Bifunctional glutathionylspermidine
FT                   synthetase/amidase"
FT                   /id="PRO_0000070444"
FT   DOMAIN          34..176
FT                   /note="Peptidase C51"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU00048"
FT   REGION          2..195
FT                   /note="Gsp amidase"
FT   REGION          196..205
FT                   /note="Linker"
FT   REGION          206..619
FT                   /note="Gsp synthetase"
FT   ACT_SITE        59
FT                   /note="S-(gamma-glutamyl-cysteinyl-glycyl)-cysteine
FT                   intermediate"
FT                   /evidence="ECO:0000250"
FT   BINDING         58
FT                   /ligand="glutathionylspermidine"
FT                   /ligand_id="ChEBI:CHEBI:57835"
FT                   /evidence="ECO:0000250"
FT   BINDING         64
FT                   /ligand="glutathionylspermidine"
FT                   /ligand_id="ChEBI:CHEBI:57835"
FT                   /evidence="ECO:0000250"
FT   BINDING         78..81
FT                   /ligand="glutathionylspermidine"
FT                   /ligand_id="ChEBI:CHEBI:57835"
FT                   /evidence="ECO:0000250"
FT   BINDING         149
FT                   /ligand="glutathionylspermidine"
FT                   /ligand_id="ChEBI:CHEBI:57835"
FT                   /evidence="ECO:0000250"
FT   BINDING         316..318
FT                   /ligand="ATP"
FT                   /ligand_id="ChEBI:CHEBI:30616"
FT                   /evidence="ECO:0000250"
FT   BINDING         316
FT                   /ligand="glutathione"
FT                   /ligand_id="ChEBI:CHEBI:57925"
FT                   /evidence="ECO:0000250"
FT   BINDING         318
FT                   /ligand="Mg(2+)"
FT                   /ligand_id="ChEBI:CHEBI:18420"
FT                   /ligand_label="1"
FT                   /evidence="ECO:0000250"
FT   BINDING         330
FT                   /ligand="Mg(2+)"
FT                   /ligand_id="ChEBI:CHEBI:18420"
FT                   /ligand_label="1"
FT                   /evidence="ECO:0000250"
FT   BINDING         330
FT                   /ligand="Mg(2+)"
FT                   /ligand_id="ChEBI:CHEBI:18420"
FT                   /ligand_label="2"
FT                   /evidence="ECO:0000250"
FT   BINDING         332
FT                   /ligand="Mg(2+)"
FT                   /ligand_id="ChEBI:CHEBI:18420"
FT                   /ligand_label="2"
FT                   /evidence="ECO:0000250"
FT   BINDING         335
FT                   /ligand="glutathione"
FT                   /ligand_id="ChEBI:CHEBI:57925"
FT                   /evidence="ECO:0000250"
FT   BINDING         391
FT                   /ligand="spermidine"
FT                   /ligand_id="ChEBI:CHEBI:57834"
FT                   /evidence="ECO:0000250"
FT   BINDING         392
FT                   /ligand="glutathione"
FT                   /ligand_id="ChEBI:CHEBI:57925"
FT                   /evidence="ECO:0000250"
FT   BINDING         446
FT                   /ligand="glutathione"
FT                   /ligand_id="ChEBI:CHEBI:57925"
FT                   /evidence="ECO:0000250"
FT   BINDING         498
FT                   /ligand="ATP"
FT                   /ligand_id="ChEBI:CHEBI:30616"
FT                   /evidence="ECO:0000250"
FT   BINDING         533
FT                   /ligand="ATP"
FT                   /ligand_id="ChEBI:CHEBI:30616"
FT                   /evidence="ECO:0000250"
FT   BINDING         539..540
FT                   /ligand="ATP"
FT                   /ligand_id="ChEBI:CHEBI:30616"
FT                   /evidence="ECO:0000250"
FT   BINDING         568..571
FT                   /ligand="ATP"
FT                   /ligand_id="ChEBI:CHEBI:30616"
FT                   /evidence="ECO:0000250"
FT   BINDING         582
FT                   /ligand="ATP"
FT                   /ligand_id="ChEBI:CHEBI:30616"
FT                   /evidence="ECO:0000250"
FT   BINDING         603..605
FT                   /ligand="ATP"
FT                   /ligand_id="ChEBI:CHEBI:30616"
FT                   /evidence="ECO:0000250"
FT   BINDING         610
FT                   /ligand="spermidine"
FT                   /ligand_id="ChEBI:CHEBI:57834"
FT                   /evidence="ECO:0000250"
FT   SITE            131
FT                   /note="Increases nucleophilicity of active site Cys; for
FT                   amidase activity"
FT                   /evidence="ECO:0000250"
FT   SITE            316
FT                   /note="Transition state stabilizer; for synthetase
FT                   activity"
FT                   /evidence="ECO:0000250"
FT   MOD_RES         59
FT                   /note="Cysteine sulfenic acid (-SOH); transient"
FT                   /evidence="ECO:0000250"
SQ   SEQUENCE   619 AA;  70532 MW;  07FB43D8A0B2933C CRC64;
     MSKGTTSQDA PFGTLLGYAP GGVAIYSSDY SSLDPQEYED DAVFRSYIDD EYMGHKWQCV
     EFARRFLFLN YGVVFTDVGM AWEIFSLRFL REVVNDNILP LQAFPNGSPR APVAGALLIW
     DKGGEFKDTG HVAIITQLHG NKVRIAEQNV IHSPLPQGQQ WTRELEMVVE NGCYTLKDTF
     DDTTILGWMI QTEDTEYSLP QPEIAGELLK ISGARLENKG QFDGKWLDEK DPLQNAYVQA
     NGQVINQDPY HYYTITESAE QELIKATNEL HLMYLHATDK VLKDDNLLAL FDIPKILWPR
     LRLSWQRRRH HMITGRMDFC MDERGLKVYE YNADSASCHT EAGLILERWA EQGYKGNGFN
     PAEGLINELA GAWKHSRARP FVHIMQDKDI EENYHAQFME QALHQAGFET RILRGLDELG
     WDAAGQLIDG EGRLVNCVWK TWAWETAFDQ IREVSDREFA AVPIRTGHPQ NEVRLIDVLL
     RPEVLVFEPL WTVIPGNKAI LPILWSLFPH HRYLLDTDFT VNDELVKTGY AVKPIAGRCG
     SNIDLVSHHE EVLDKTSGKF AEQKNIYQQL WCLPKVDGKY IQVCTFTVGG NYGGTCLRGD
     ESLVIKKESD IEPLIVVKK
//