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Protein

Bifunctional glutathionylspermidine synthetase/amidase

Gene

gss

Organism
Escherichia coli (strain K12)
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

Catalyzes the formation of an amide bond between glutathione (GSH) and spermidine coupled with hydrolysis of ATP; also catalyzes the opposing reaction, i.e. the hydrolysis of glutathionylspermidine (Gsp) back to glutathione and spermidine. The amidase active site can also hydrolyze Gsp-disulfide (Gsp-S-S-Gsp) to Gsp-SG and Gsp S-thiolated proteins (GspSSPs) to GSH S-thiolated protein (GSSPs). Likely acts synergistically with glutaredoxin to regulate the redox environment of E.coli and defend against oxidative damage. In vitro, the amidase active site also catalyzes hydrolysis of amide and ester derivatives of glutathione (e.g. glutathione ethyl ester and glutathione amide) but lacks activity toward acetylspermidine (N1 and N8) and acetylspermine (N1).4 Publications

Catalytic activityi

Glutathione + spermidine + ATP = glutathionylspermidine + ADP + phosphate.
Glutathionylspermidine + H2O = glutathione + spermidine.

Enzyme regulationi

When exposed to oxidative stress, Gsp amidase activity is transiently inhibited in vivo by oxidation of the catalytic Cys-59 thiol to sulfenic acid; this modification does not affect Gsp synthetase activity. Gsp amidase activity is negatively autoregulated by the Gsp synthetase domain, and is activated by the Gsp synthetase substrates, GSH and ATP-Mg2+; the occupancy of the synthetase active site may initiate communication through the protein as manifest by the release of inhibition of the amidase activity. A tetrahedral phosphonate analog of glutathionylspermidine, designed as a mimic of the proposed tetrahedral intermediate for either reaction, inhibits the synthetase activity (Ki of 10 µM) but does not inhibit the amidase activity. Amidase activity is inhibited by iodoacetamide in vitro.3 Publications

Kineticsi

kcat is 7 sec(-1) for Gsp synthetase activity at pH 6.8 and 2.1 sec(-1) for Gsp amidase activity at pH 7.5.1 Publication

  1. KM=100 µM for ATP (at pH 6.8)2 Publications
  2. KM=800 µM for glutathione (at pH 6.8)2 Publications
  3. KM=218 µM for glutathione2 Publications
  4. KM=60 µM for spermidine (at pH 6.8)2 Publications
  5. KM=20 µM for spermidine (at pH 7.5)2 Publications
  6. KM=76 µM for spermidine2 Publications
  7. KM=900 µM for glutathionylspermidine (at pH 7.5)2 Publications

    pH dependencei

    Optimum pH is around 6.8 for Gsp synthetase activity.1 Publication

    Pathway:iglutathione metabolism

    This protein is involved in the pathway glutathione metabolism, which is part of Sulfur metabolism.
    View all proteins of this organism that are known to be involved in the pathway glutathione metabolism and in Sulfur metabolism.

    Pathway:ispermidine metabolism

    This protein is involved in the pathway spermidine metabolism, which is part of Amine and polyamine metabolism.
    View all proteins of this organism that are known to be involved in the pathway spermidine metabolism and in Amine and polyamine metabolism.

    Sites

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Binding sitei58 – 581Gsp
    Active sitei59 – 591S-(gamma-glutamyl-cysteinyl-glycyl)-cysteine intermediate1 Publication
    Binding sitei64 – 641Gsp
    Sitei131 – 1311Increases nucleophilicity of active site Cys; for amidase activity
    Binding sitei149 – 1491Gsp
    Binding sitei316 – 3161Glutathione
    Sitei316 – 3161Transition state stabilizer; for synthetase activity
    Metal bindingi318 – 3181Magnesium 1
    Metal bindingi330 – 3301Magnesium 1
    Metal bindingi330 – 3301Magnesium 2
    Metal bindingi332 – 3321Magnesium 2
    Binding sitei335 – 3351Glutathione
    Binding sitei391 – 3911Spermidine
    Binding sitei392 – 3921Glutathione
    Binding sitei446 – 4461Glutathione
    Binding sitei498 – 4981ATP
    Binding sitei533 – 5331ATP
    Binding sitei582 – 5821ATP
    Binding sitei610 – 6101Spermidine

    Regions

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Nucleotide bindingi316 – 3183ATP
    Nucleotide bindingi539 – 5402ATP
    Nucleotide bindingi568 – 5714ATP
    Nucleotide bindingi603 – 6053ATP

    GO - Molecular functioni

    • ATP binding Source: UniProtKB-KW
    • glutathionylspermidine amidase activity Source: EcoCyc
    • glutathionylspermidine synthase activity Source: EcoCyc
    • metal ion binding Source: UniProtKB-KW

    GO - Biological processi

    Complete GO annotation...

    Keywords - Molecular functioni

    Hydrolase, Ligase

    Keywords - Ligandi

    ATP-binding, Magnesium, Metal-binding, Nucleotide-binding

    Enzyme and pathway databases

    BioCyciEcoCyc:GSP-MONOMER.
    ECOL316407:JW2956-MONOMER.
    MetaCyc:GSP-MONOMER.
    BRENDAi3.5.1.78. 2026.
    6.3.1.8. 2026.
    UniPathwayiUPA00204.
    UPA00819.

    Protein family/group databases

    MEROPSiC51.A01.

    Names & Taxonomyi

    Protein namesi
    Recommended name:
    Bifunctional glutathionylspermidine synthetase/amidase
    Short name:
    GspSA
    Including the following 2 domains:
    Glutathionylspermidine amidase (EC:3.5.1.78)
    Short name:
    Gsp amidase
    Alternative name(s):
    Glutathionylspermidine amidohydrolase [spermidine-forming]
    Glutathionylspermidine synthetase (EC:6.3.1.8)
    Short name:
    Gsp synthetase
    Alternative name(s):
    Glutathione:spermidine ligase [ADP-forming]
    Gsp synthase
    Gene namesi
    Name:gss
    Synonyms:gsp
    Ordered Locus Names:b2988, JW2956
    OrganismiEscherichia coli (strain K12)
    Taxonomic identifieri83333 [NCBI]
    Taxonomic lineageiBacteriaProteobacteriaGammaproteobacteriaEnterobacterialesEnterobacteriaceaeEscherichia
    ProteomesiUP000000318 Componenti: Chromosome UP000000625 Componenti: Chromosome

    Organism-specific databases

    EcoGeneiEG12882. gss.

    Pathology & Biotechi

    Disruption phenotypei

    Cells lacking this gene do not produce Gsp under anaerobic conditions. Cells lacking both this gene and glutaredoxin (grxA or grxB) become hypersensitive to H2O2; they are even more susceptible to oxidative damage than the single mutant lacking glutaredoxin only.2 Publications

    Mutagenesis

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Mutagenesisi59 – 591C → A: Loss of amidase activity. 1 Publication
    Mutagenesisi173 – 1731C → A: No effect on amidase activity. 1 Publication
    Mutagenesisi316 – 3161R → E: Loss of synthetase activity.
    Mutagenesisi335 – 3351S → A: 3.6-fold decrease in GSH affinity, 1.6-fold decrease in spermidine activity, and 1.3-fold decrease in synthetase activity. 1 Publication
    Mutagenesisi337 – 3371S → A: No effect on GSH and spermidine affinity, but 2-fold decrease in synthetase activity. 1 Publication
    Mutagenesisi338 – 3381C → A: 10-fold decrease in GSH affinity, 5-fold decrease in spermidine activity, but no effect on synthetase activity. 1 Publication
    Mutagenesisi391 – 3911E → A: 2-fold decrease in GSH affinity, 60-fold decrease in spermidine activity, and 10-fold decrease in synthetase activity. 1 Publication
    Mutagenesisi392 – 3921E → A: 33-fold decrease in GSH affinity, 13-fold decrease in spermidine activity, and 6-fold decrease in synthetase activity. 1 Publication
    Mutagenesisi441 – 4411T → A: 3-fold decrease in GSH affinity, 21-fold decrease in spermidine activity, and 17-fold decrease in synthetase activity. 1 Publication
    Mutagenesisi538 – 5381R → A: 6-fold decrease in GSH affinity, 2.4-fold decrease in spermidine activity, and 4-fold decrease in synthetase activity. 1 Publication
    Mutagenesisi598 – 5981R → A: 10-fold increase in GSH affinity, 9-fold decrease in spermidine activity, and 15-fold decrease in synthetase activity. 1 Publication

    PTM / Processingi

    Molecule processing

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Initiator methioninei1 – 11Removed1 Publication
    Chaini2 – 619618Bifunctional glutathionylspermidine synthetase/amidasePRO_0000070443Add
    BLAST

    Amino acid modifications

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Modified residuei59 – 591Cysteine sulfenic acid (-SOH); transient1 Publication

    Post-translational modificationi

    Oxidation of Cys-59 to sulfenic acid during oxidative stress selectively inhibits the amidase activity which leads to a rapid increase in the amounts of intracellular Gsp and Gsp S-thiolated proteins (GspSSPs).1 Publication

    Keywords - PTMi

    Oxidation

    Proteomic databases

    PaxDbiP0AES0.
    PRIDEiP0AES0.

    Expressioni

    Inductioni

    Expression level is unaffected by H2O2; however Gsp rapidly accumulates in E.coli in the presence of H2O2.

    Interactioni

    Subunit structurei

    Homodimer.2 Publications

    Binary interactionsi

    WithEntry#Exp.IntActNotes
    fixBP315742EBI-557080,EBI-554030
    rhoP0AG304EBI-557080,EBI-545468

    Protein-protein interaction databases

    DIPiDIP-36018N.
    IntActiP0AES0. 14 interactions.
    MINTiMINT-1234290.
    STRINGi511145.b2988.

    Structurei

    Secondary structure

    1
    619
    Legend: HelixTurnBeta strand
    Show more details
    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Beta strandi15 – 195Combined sources
    Turni20 – 223Combined sources
    Beta strandi23 – 264Combined sources
    Helixi35 – 406Combined sources
    Helixi42 – 443Combined sources
    Beta strandi45 – 484Combined sources
    Beta strandi51 – 555Combined sources
    Helixi59 – 7113Combined sources
    Beta strandi72 – 743Combined sources
    Helixi81 – 866Combined sources
    Beta strandi89 – 924Combined sources
    Turni93 – 953Combined sources
    Beta strandi98 – 1003Combined sources
    Beta strandi102 – 1054Combined sources
    Beta strandi108 – 1103Combined sources
    Beta strandi117 – 1204Combined sources
    Helixi124 – 1263Combined sources
    Turni127 – 1293Combined sources
    Beta strandi131 – 1388Combined sources
    Beta strandi140 – 1467Combined sources
    Beta strandi148 – 1503Combined sources
    Beta strandi162 – 1709Combined sources
    Beta strandi173 – 1775Combined sources
    Beta strandi179 – 1824Combined sources
    Beta strandi185 – 1928Combined sources
    Helixi206 – 2094Combined sources
    Beta strandi212 – 2154Combined sources
    Beta strandi229 – 2313Combined sources
    Helixi232 – 24110Combined sources
    Beta strandi245 – 2473Combined sources
    Beta strandi251 – 2566Combined sources
    Helixi257 – 28327Combined sources
    Helixi285 – 2884Combined sources
    Helixi289 – 2913Combined sources
    Helixi295 – 2973Combined sources
    Helixi298 – 30710Combined sources
    Helixi309 – 3113Combined sources
    Beta strandi314 – 3229Combined sources
    Beta strandi325 – 3328Combined sources
    Helixi339 – 3435Combined sources
    Helixi345 – 3539Combined sources
    Turni361 – 3644Combined sources
    Helixi365 – 37410Combined sources
    Beta strandi380 – 3867Combined sources
    Helixi390 – 40516Combined sources
    Beta strandi409 – 4168Combined sources
    Beta strandi423 – 4253Combined sources
    Beta strandi437 – 4426Combined sources
    Helixi444 – 45310Combined sources
    Turni456 – 4583Combined sources
    Beta strandi459 – 4613Combined sources
    Helixi475 – 4795Combined sources
    Beta strandi485 – 4884Combined sources
    Helixi490 – 4934Combined sources
    Turni494 – 4963Combined sources
    Helixi500 – 5078Combined sources
    Beta strandi517 – 5204Combined sources
    Helixi523 – 5286Combined sources
    Beta strandi530 – 5345Combined sources
    Turni539 – 5424Combined sources
    Beta strandi544 – 5463Combined sources
    Beta strandi552 – 5554Combined sources
    Turni559 – 5624Combined sources
    Beta strandi565 – 5695Combined sources
    Beta strandi579 – 58810Combined sources
    Beta strandi591 – 60414Combined sources
    Beta strandi609 – 6124Combined sources
    Beta strandi614 – 6174Combined sources

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    2IO7X-ray2.70A/B1-619[»]
    2IO8X-ray2.10A/B1-619[»]
    2IO9X-ray2.20A/B1-619[»]
    2IOAX-ray2.80A/B1-619[»]
    2IOBX-ray2.20A/B1-619[»]
    3A2YX-ray1.95A1-197[»]
    3A2ZX-ray1.50A1-197[»]
    3A30X-ray2.20A1-197[»]
    3O98X-ray2.80A/B1-619[»]
    ProteinModelPortaliP0AES0.
    SMRiP0AES0. Positions 8-618.
    ModBaseiSearch...
    MobiDBiSearch...

    Miscellaneous databases

    EvolutionaryTraceiP0AES0.

    Family & Domainsi

    Domains and Repeats

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Domaini34 – 176143Peptidase C51PROSITE-ProRule annotationAdd
    BLAST

    Region

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Regioni2 – 195194Gsp amidaseAdd
    BLAST
    Regioni78 – 814Gsp binding
    Regioni196 – 20510Linker
    Regioni206 – 619414Gsp synthetaseAdd
    BLAST

    Domaini

    The two activities reside in distinct domains (N-terminal amidase and C-terminal synthetase). The two domains expressed independently are folded and functional; liberation of the amidase domain from the synthetase domain highly activates the amidase activity.2 Publications

    Sequence similaritiesi

    In the C-terminal section; belongs to the glutathionylspermidine synthase preATP-grasp family.Curated
    Contains 1 peptidase C51 domain.PROSITE-ProRule annotation

    Phylogenomic databases

    eggNOGiCOG0754.
    HOGENOMiHOG000124980.
    InParanoidiP0AES0.
    KOiK01460.
    OMAiYMGYKWQ.
    OrthoDBiEOG66XB93.

    Family and domain databases

    InterProiIPR007921. CHAP_dom.
    IPR005494. GSPS_pre-ATP-grasp-like_dom.
    IPR016185. PreATP-grasp_dom.
    [Graphical view]
    PfamiPF05257. CHAP. 1 hit.
    PF03738. GSP_synth. 1 hit.
    [Graphical view]
    SUPFAMiSSF52440. SSF52440. 1 hit.
    PROSITEiPS50911. CHAP. 1 hit.
    [Graphical view]

    Sequencei

    Sequence statusi: Complete.

    Sequence processingi: The displayed sequence is further processed into a mature form.

    P0AES0-1 [UniParc]FASTAAdd to basket

    « Hide

            10         20         30         40         50
    MSKGTTSQDA PFGTLLGYAP GGVAIYSSDY SSLDPQEYED DAVFRSYIDD
    60 70 80 90 100
    EYMGHKWQCV EFARRFLFLN YGVVFTDVGM AWEIFSLRFL REVVNDNILP
    110 120 130 140 150
    LQAFPNGSPR APVAGALLIW DKGGEFKDTG HVAIITQLHG NKVRIAEQNV
    160 170 180 190 200
    IHSPLPQGQQ WTRELEMVVE NGCYTLKDTF DDTTILGWMI QTEDTEYSLP
    210 220 230 240 250
    QPEIAGELLK ISGARLENKG QFDGKWLDEK DPLQNAYVQA NGQVINQDPY
    260 270 280 290 300
    HYYTITESAE QELIKATNEL HLMYLHATDK VLKDDNLLAL FDIPKILWPR
    310 320 330 340 350
    LRLSWQRRRH HMITGRMDFC MDERGLKVYE YNADSASCHT EAGLILERWA
    360 370 380 390 400
    EQGYKGNGFN PAEGLINELA GAWKHSRARP FVHIMQDKDI EENYHAQFME
    410 420 430 440 450
    QALHQAGFET RILRGLDELG WDAAGQLIDG EGRLVNCVWK TWAWETAFDQ
    460 470 480 490 500
    IREVSDREFA AVPIRTGHPQ NEVRLIDVLL RPEVLVFEPL WTVIPGNKAI
    510 520 530 540 550
    LPILWSLFPH HRYLLDTDFT VNDELVKTGY AVKPIAGRCG SNIDLVSHHE
    560 570 580 590 600
    EVLDKTSGKF AEQKNIYQQL WCLPKVDGKY IQVCTFTVGG NYGGTCLRGD
    610
    ESLVIKKESD IEPLIVVKK
    Length:619
    Mass (Da):70,532
    Last modified:December 20, 2005 - v1
    Checksum:i07FB43D8A0B2933C
    GO

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    U23148 Genomic DNA. Translation: AAC43339.1.
    U28377 Genomic DNA. Translation: AAA69155.1.
    U00096 Genomic DNA. Translation: AAC76024.1.
    AP009048 Genomic DNA. Translation: BAE77049.1.
    PIRiA57538.
    RefSeqiNP_417462.1. NC_000913.3.
    WP_001297309.1. NZ_CP010445.1.

    Genome annotation databases

    EnsemblBacteriaiAAC76024; AAC76024; b2988.
    BAE77049; BAE77049; BAE77049.
    GeneIDi947474.
    KEGGieco:b2988.
    PATRICi32121392. VBIEscCol129921_3083.

    Cross-referencesi

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    U23148 Genomic DNA. Translation: AAC43339.1.
    U28377 Genomic DNA. Translation: AAA69155.1.
    U00096 Genomic DNA. Translation: AAC76024.1.
    AP009048 Genomic DNA. Translation: BAE77049.1.
    PIRiA57538.
    RefSeqiNP_417462.1. NC_000913.3.
    WP_001297309.1. NZ_CP010445.1.

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    2IO7X-ray2.70A/B1-619[»]
    2IO8X-ray2.10A/B1-619[»]
    2IO9X-ray2.20A/B1-619[»]
    2IOAX-ray2.80A/B1-619[»]
    2IOBX-ray2.20A/B1-619[»]
    3A2YX-ray1.95A1-197[»]
    3A2ZX-ray1.50A1-197[»]
    3A30X-ray2.20A1-197[»]
    3O98X-ray2.80A/B1-619[»]
    ProteinModelPortaliP0AES0.
    SMRiP0AES0. Positions 8-618.
    ModBaseiSearch...
    MobiDBiSearch...

    Protein-protein interaction databases

    DIPiDIP-36018N.
    IntActiP0AES0. 14 interactions.
    MINTiMINT-1234290.
    STRINGi511145.b2988.

    Protein family/group databases

    MEROPSiC51.A01.

    Proteomic databases

    PaxDbiP0AES0.
    PRIDEiP0AES0.

    Protocols and materials databases

    Structural Biology KnowledgebaseSearch...

    Genome annotation databases

    EnsemblBacteriaiAAC76024; AAC76024; b2988.
    BAE77049; BAE77049; BAE77049.
    GeneIDi947474.
    KEGGieco:b2988.
    PATRICi32121392. VBIEscCol129921_3083.

    Organism-specific databases

    EchoBASEiEB2720.
    EcoGeneiEG12882. gss.

    Phylogenomic databases

    eggNOGiCOG0754.
    HOGENOMiHOG000124980.
    InParanoidiP0AES0.
    KOiK01460.
    OMAiYMGYKWQ.
    OrthoDBiEOG66XB93.

    Enzyme and pathway databases

    UniPathwayiUPA00204.
    UPA00819.
    BioCyciEcoCyc:GSP-MONOMER.
    ECOL316407:JW2956-MONOMER.
    MetaCyc:GSP-MONOMER.
    BRENDAi3.5.1.78. 2026.
    6.3.1.8. 2026.

    Miscellaneous databases

    EvolutionaryTraceiP0AES0.
    PROiP0AES0.

    Family and domain databases

    InterProiIPR007921. CHAP_dom.
    IPR005494. GSPS_pre-ATP-grasp-like_dom.
    IPR016185. PreATP-grasp_dom.
    [Graphical view]
    PfamiPF05257. CHAP. 1 hit.
    PF03738. GSP_synth. 1 hit.
    [Graphical view]
    SUPFAMiSSF52440. SSF52440. 1 hit.
    PROSITEiPS50911. CHAP. 1 hit.
    [Graphical view]
    ProtoNetiSearch...

    Publicationsi

    « Hide 'large scale' publications
    1. "Glutathionylspermidine metabolism in Escherichia coli. Purification, cloning, overproduction, and characterization of a bifunctional glutathionylspermidine synthetase/amidase."
      Bollinger J.M. Jr., Kwon D.S., Huisman G.W., Kolter R., Walsh C.T.
      J. Biol. Chem. 270:14031-14041(1995) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-7, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT.
      Strain: B and K12.
    2. Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
      Strain: K12 / MG1655 / ATCC 47076.
    3. "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.
    4. "Evidence for a glutathionyl-enzyme intermediate in the amidase activity of the bifunctional glutathionylspermidine synthetase/amidase from Escherichia coli."
      Lin C.H., Kwon D.S., Bollinger J.M. Jr., Walsh C.T.
      Biochemistry 36:14930-14938(1997) [PubMed] [Europe PMC] [Abstract]
      Cited for: REACTION MECHANISM OF AMIDASE ACTIVITY, ACTIVE SITE AT CYS-59, ENZYME REGULATION, MUTAGENESIS OF CYS-59 AND CYS-173.
    5. "Dissection of glutathionylspermidine synthetase/amidase from Escherichia coli into autonomously folding and functional synthetase and amidase domains."
      Kwon D.S., Lin C.H., Chen S., Coward J.K., Walsh C.T., Bollinger J.M. Jr.
      J. Biol. Chem. 272:2429-2436(1997) [PubMed] [Europe PMC] [Abstract]
      Cited for: FUNCTION, CATALYTIC ACTIVITY, DOMAIN, SUBSTRATE SPECIFICITY, ENZYME REGULATION.
    6. "Comparison of the functions of glutathionylspermidine synthetase/amidase from E. coli and its predicted homologues YgiC and YjfC."
      Sui L., Warren J.C., Russell J.P., Stourman N.V.
      Int. J. Biochem. Mol. Biol. 3:302-312(2012) [PubMed] [Europe PMC] [Abstract]
      Cited for: FUNCTION, CATALYTIC ACTIVITY, DISRUPTION PHENOTYPE.
      Strain: K12.
    7. "Dual binding sites for translocation catalysis by Escherichia coli glutathionylspermidine synthetase."
      Pai C.H., Chiang B.Y., Ko T.P., Chou C.C., Chong C.M., Yen F.J., Chen S., Coward J.K., Wang A.H., Lin C.H.
      EMBO J. 25:5970-5982(2006) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) OF APOENZYME AND IN COMPLEXES WITH SUBSTRATE; PRODUCT AND INHIBITOR, DOMAIN BOUNDARIES, SUBUNIT, REACTION MECHANISM OF SYNTHETASE ACTIVITY, KINETIC PARAMETERS, SITE AT ARG-316, MUTAGENESIS OF SER-335; SER-337; CYS-338; GLU-391; GLU-392; THR-441; ARG-538 AND ARG-598.
    8. "Protein S-thiolation by glutathionylspermidine (Gsp): the role of Escherichia coli Gsp synthetase/amidase in redox regulation."
      Chiang B.Y., Chen T.C., Pai C.H., Chou C.C., Chen H.H., Ko T.P., Hsu W.H., Chang C.Y., Wu W.F., Wang A.H., Lin C.H.
      J. Biol. Chem. 285:25345-25353(2010) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (1.50 ANGSTROMS) OF 1-197, FUNCTION, ROLE IN REDOX REGULATION, CATALYTIC ACTIVITY, ENZYME REGULATION, OXIDATION AT CYS-59, DISRUPTION PHENOTYPE.
    9. "Structure and mechanism of Escherichia coli glutathionylspermidine amidase belonging to the family of cysteine; histidine-dependent amidohydrolases/peptidases."
      Pai C.H., Wu H.J., Lin C.H., Wang A.H.
      Protein Sci. 20:557-566(2011) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (1.95 ANGSTROMS) OF MUTANT ALA-59 IN COMPLEXES WITH ADP; GLUTATHIONYLSPERMIDINE AND MAGNESIUM, ACTIVE SITES, CATALYTIC MECHANISM OF AMIDASE ACTIVITY.

    Entry informationi

    Entry nameiGSP_ECOLI
    AccessioniPrimary (citable) accession number: P0AES0
    Secondary accession number(s): P43675, Q2M9K7
    Entry historyi
    Integrated into UniProtKB/Swiss-Prot: December 20, 2005
    Last sequence update: December 20, 2005
    Last modified: July 22, 2015
    This is version 86 of the entry and version 1 of the sequence. [Complete history]
    Entry statusiReviewed (UniProtKB/Swiss-Prot)
    Annotation programProkaryotic Protein Annotation Program

    Miscellaneousi

    Miscellaneous

    Gsp forms mixed disulfides with the thiols of a variety of E.coli proteins. These mixed disulfides represent a previously uncharacterized type of post-translational modification. The level of these proteins is increased by oxidative stress, which implies that Gsp might protect protein thiols against irreversible oxidation (PubMed:20530482).1 Publication
    No metal ion is required for the amidase activity.1 Publication
    Gsp hydrolysis to GSH and spermidine proceeds with formation of a glutathionyl acyl-enzyme intermediate, utilizing a cysteine residue as the catalytic nucleophile (PubMed:9398217). For Gsp synthesis, GSH is likely phosphorylated at one of two GSH-binding sites to form an acylphosphate intermediate that then translocates to the other site for subsequent nucleophilic addition of spermidine (PubMed:17124497).2 Publications

    Keywords - Technical termi

    3D-structure, Complete proteome, Direct protein sequencing, Multifunctional enzyme, Reference proteome

    Documents

    1. Escherichia coli
      Escherichia coli (strain K12): entries and cross-references to EcoGene
    2. PATHWAY comments
      Index of metabolic and biosynthesis pathways
    3. PDB cross-references
      Index of Protein Data Bank (PDB) cross-references
    4. Peptidase families
      Classification of peptidase families and list of entries
    5. SIMILARITY comments
      Index of protein domains and families

    External Data

    Dasty 3

    Similar proteinsi

    Links to similar proteins from the UniProt Reference Clusters (UniRef) at 100%, 90% and 50% sequence identity:
    100%UniRef100 combines identical sequences and sub-fragments with 11 or more residues from any organism into Uniref entry.
    90%UniRef90 is built by clustering UniRef100 sequences that have at least 90% sequence identity to, and 80% overlap with, the longest sequence (a.k.a seed sequence).
    50%UniRef50 is built by clustering UniRef90 seed sequences that have at least 50% sequence identity to, and 80% overlap with, the longest sequence in the cluster.