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P0AES0

- GSP_ECOLI

UniProt

P0AES0 - GSP_ECOLI

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Protein

Bifunctional glutathionylspermidine synthetase/amidase

Gene

gss

Organism
Escherichia coli (strain K12)
Status
Reviewed - 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

Pathwayi

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

  1. ATP binding Source: UniProtKB-KW
  2. glutathionylspermidine amidase activity Source: EcoCyc
  3. glutathionylspermidine synthase activity Source: EcoCyc
  4. metal ion binding Source: UniProtKB-KW

GO - Biological processi

  1. glutathione metabolic process Source: UniProtKB-UniPathway
  2. spermidine metabolic process Source: UniProtKB-UniPathway
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.
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: Chromosome, UP000000625: 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.

Gene expression databases

GenevestigatoriP0AES0.

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

3D structure databases

Select the link destinations:
PDBe
RCSB PDB
PDBj
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 [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:
EMBL
GenBank
DDBJ
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.
YP_491185.1. NC_007779.1.

Genome annotation databases

EnsemblBacteriaiAAC76024; AAC76024; b2988.
BAE77049; BAE77049; BAE77049.
GeneIDi12932186.
947474.
KEGGiecj:Y75_p2917.
eco:b2988.
PATRICi32121392. VBIEscCol129921_3083.

Cross-referencesi

Sequence databases

Select the link destinations:
EMBL
GenBank
DDBJ
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 .
PIRi A57538.
RefSeqi NP_417462.1. NC_000913.3.
YP_491185.1. NC_007779.1.

3D structure databases

Select the link destinations:
PDBe
RCSB PDB
PDBj
Links Updated
Entry Method Resolution (Å) Chain Positions PDBsum
2IO7 X-ray 2.70 A/B 1-619 [» ]
2IO8 X-ray 2.10 A/B 1-619 [» ]
2IO9 X-ray 2.20 A/B 1-619 [» ]
2IOA X-ray 2.80 A/B 1-619 [» ]
2IOB X-ray 2.20 A/B 1-619 [» ]
3A2Y X-ray 1.95 A 1-197 [» ]
3A2Z X-ray 1.50 A 1-197 [» ]
3A30 X-ray 2.20 A 1-197 [» ]
3O98 X-ray 2.80 A/B 1-619 [» ]
ProteinModelPortali P0AES0.
SMRi P0AES0. Positions 8-618.
ModBasei Search...
MobiDBi Search...

Protein-protein interaction databases

DIPi DIP-36018N.
IntActi P0AES0. 14 interactions.
MINTi MINT-1234290.
STRINGi 511145.b2988.

Protein family/group databases

MEROPSi C51.A01.

Proteomic databases

PaxDbi P0AES0.
PRIDEi P0AES0.

Protocols and materials databases

Structural Biology Knowledgebase Search...

Genome annotation databases

EnsemblBacteriai AAC76024 ; AAC76024 ; b2988 .
BAE77049 ; BAE77049 ; BAE77049 .
GeneIDi 12932186.
947474.
KEGGi ecj:Y75_p2917.
eco:b2988.
PATRICi 32121392. VBIEscCol129921_3083.

Organism-specific databases

EchoBASEi EB2720.
EcoGenei EG12882. gss.

Phylogenomic databases

eggNOGi COG0754.
HOGENOMi HOG000124980.
InParanoidi P0AES0.
KOi K01460.
OMAi YMGYKWQ.
OrthoDBi EOG66XB93.

Enzyme and pathway databases

UniPathwayi UPA00204 .
UPA00819 .
BioCyci EcoCyc:GSP-MONOMER.
ECOL316407:JW2956-MONOMER.
MetaCyc:GSP-MONOMER.

Miscellaneous databases

EvolutionaryTracei P0AES0.
PROi P0AES0.

Gene expression databases

Genevestigatori P0AES0.

Family and domain databases

InterProi IPR007921. CHAP_dom.
IPR005494. GSPS_pre-ATP-grasp-like_dom.
IPR016185. PreATP-grasp_dom.
[Graphical view ]
Pfami PF05257. CHAP. 1 hit.
PF03738. GSP_synth. 1 hit.
[Graphical view ]
SUPFAMi SSF52440. SSF52440. 1 hit.
PROSITEi PS50911. CHAP. 1 hit.
[Graphical view ]
ProtoNeti Search...

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: October 29, 2014
This is version 80 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