GSP_ECOLI
ID GSP_ECOLI Reviewed; 619 AA.
AC P0AES0; P43675; Q2M9K7;
DT 20-DEC-2005, integrated into UniProtKB/Swiss-Prot.
DT 20-DEC-2005, sequence version 1.
DT 03-AUG-2022, entry version 125.
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:0000269|PubMed:20530482, ECO:0000269|PubMed:23097746, ECO:0000269|PubMed:7775463, ECO:0000269|PubMed:8999955};
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:0000269|PubMed:20530482, ECO:0000269|PubMed:23097746, ECO:0000269|PubMed:7775463, ECO:0000269|PubMed:8999955};
DE AltName: Full=Glutathione:spermidine ligase [ADP-forming];
DE AltName: Full=Gsp synthase;
GN Name=gss; Synonyms=gsp; OrderedLocusNames=b2988, JW2956;
OS Escherichia coli (strain K12).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=83333;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-7, FUNCTION,
RP CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND SUBUNIT.
RC STRAIN=B, and K12;
RX PubMed=7775463; DOI=10.1074/jbc.270.23.14031;
RA Bollinger J.M. Jr., Kwon D.S., Huisman G.W., Kolter R., Walsh C.T.;
RT "Glutathionylspermidine metabolism in Escherichia coli. Purification,
RT cloning, overproduction, and characterization of a bifunctional
RT glutathionylspermidine synthetase/amidase.";
RL J. Biol. Chem. 270:14031-14041(1995).
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 REACTION MECHANISM OF AMIDASE ACTIVITY, ACTIVE SITE AT CYS-59, ACTIVITY
RP REGULATION, AND MUTAGENESIS OF CYS-59 AND CYS-173.
RX PubMed=9398217; DOI=10.1021/bi9714464;
RA Lin C.H., Kwon D.S., Bollinger J.M. Jr., Walsh C.T.;
RT "Evidence for a glutathionyl-enzyme intermediate in the amidase activity of
RT the bifunctional glutathionylspermidine synthetase/amidase from Escherichia
RT coli.";
RL Biochemistry 36:14930-14938(1997).
RN [5]
RP FUNCTION, CATALYTIC ACTIVITY, DOMAIN, SUBSTRATE SPECIFICITY, AND ACTIVITY
RP REGULATION.
RX PubMed=8999955; DOI=10.1074/jbc.272.4.2429;
RA Kwon D.S., Lin C.H., Chen S., Coward J.K., Walsh C.T., Bollinger J.M. Jr.;
RT "Dissection of glutathionylspermidine synthetase/amidase from Escherichia
RT coli into autonomously folding and functional synthetase and amidase
RT domains.";
RL J. Biol. Chem. 272:2429-2436(1997).
RN [6]
RP FUNCTION, CATALYTIC ACTIVITY, AND DISRUPTION PHENOTYPE.
RC STRAIN=K12;
RX PubMed=23097746;
RA Sui L., Warren J.C., Russell J.P., Stourman N.V.;
RT "Comparison of the functions of glutathionylspermidine synthetase/amidase
RT from E. coli and its predicted homologues YgiC and YjfC.";
RL Int. J. Biochem. Mol. Biol. 3:302-312(2012).
RN [7]
RP X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) OF APOENZYME AND IN COMPLEXES WITH
RP SUBSTRATE; PRODUCT AND INHIBITOR, DOMAIN BOUNDARIES, SUBUNIT, REACTION
RP MECHANISM OF SYNTHETASE ACTIVITY, KINETIC PARAMETERS, SITE AT ARG-316, AND
RP MUTAGENESIS OF SER-335; SER-337; CYS-338; GLU-391; GLU-392; THR-441;
RP ARG-538 AND ARG-598.
RX PubMed=17124497; DOI=10.1038/sj.emboj.7601440;
RA Pai C.H., Chiang B.Y., Ko T.P., Chou C.C., Chong C.M., Yen F.J., Chen S.,
RA Coward J.K., Wang A.H., Lin C.H.;
RT "Dual binding sites for translocation catalysis by Escherichia coli
RT glutathionylspermidine synthetase.";
RL EMBO J. 25:5970-5982(2006).
RN [8]
RP X-RAY CRYSTALLOGRAPHY (1.50 ANGSTROMS) OF 1-197, FUNCTION, ROLE IN REDOX
RP REGULATION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, OXIDATION AT CYS-59,
RP AND DISRUPTION PHENOTYPE.
RX PubMed=20530482; DOI=10.1074/jbc.m110.133363;
RA Chiang B.Y., Chen T.C., Pai C.H., Chou C.C., Chen H.H., Ko T.P., Hsu W.H.,
RA Chang C.Y., Wu W.F., Wang A.H., Lin C.H.;
RT "Protein S-thiolation by glutathionylspermidine (Gsp): the role of
RT Escherichia coli Gsp synthetase/amidase in redox regulation.";
RL J. Biol. Chem. 285:25345-25353(2010).
RN [9]
RP X-RAY CRYSTALLOGRAPHY (1.95 ANGSTROMS) OF MUTANT ALA-59 IN COMPLEXES WITH
RP ADP; GLUTATHIONYLSPERMIDINE AND MAGNESIUM, ACTIVE SITES, AND CATALYTIC
RP MECHANISM OF AMIDASE ACTIVITY.
RX PubMed=21226054; DOI=10.1002/pro.589;
RA Pai C.H., Wu H.J., Lin C.H., Wang A.H.;
RT "Structure and mechanism of Escherichia coli glutathionylspermidine amidase
RT belonging to the family of cysteine; histidine-dependent
RT amidohydrolases/peptidases.";
RL Protein Sci. 20:557-566(2011).
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. The amidase active site can also
CC hydrolyze Gsp-disulfide (Gsp-S-S-Gsp) to Gsp-SG and Gsp S-thiolated
CC proteins (GspSSPs) to GSH S-thiolated protein (GSSPs). Likely acts
CC synergistically with glutaredoxin to regulate the redox environment of
CC E.coli and defend against oxidative damage. In vitro, the amidase
CC active site also catalyzes hydrolysis of amide and ester derivatives of
CC glutathione (e.g. glutathione ethyl ester and glutathione amide) but
CC lacks activity toward acetylspermidine (N1 and N8) and acetylspermine
CC (N1). {ECO:0000269|PubMed:20530482, ECO:0000269|PubMed:23097746,
CC ECO:0000269|PubMed:7775463, ECO:0000269|PubMed:8999955}.
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:0000269|PubMed:20530482, ECO:0000269|PubMed:23097746,
CC ECO:0000269|PubMed:7775463, ECO:0000269|PubMed:8999955};
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:0000269|PubMed:20530482, ECO:0000269|PubMed:23097746,
CC ECO:0000269|PubMed:7775463, ECO:0000269|PubMed:8999955};
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. Gsp amidase activity is negatively autoregulated
CC by the Gsp synthetase domain, and is activated by the Gsp synthetase
CC substrates, GSH and ATP-Mg(2+); the occupancy of the synthetase active
CC site may initiate communication through the protein as manifest by the
CC release of inhibition of the amidase activity. A tetrahedral
CC phosphonate analog of glutathionylspermidine, designed as a mimic of
CC the proposed tetrahedral intermediate for either reaction, inhibits the
CC synthetase activity (Ki of 10 uM) but does not inhibit the amidase
CC activity. Amidase activity is inhibited by iodoacetamide in vitro.
CC {ECO:0000269|PubMed:20530482, ECO:0000269|PubMed:8999955,
CC ECO:0000269|PubMed:9398217}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=100 uM for ATP (at pH 6.8) {ECO:0000269|PubMed:17124497,
CC ECO:0000269|PubMed:7775463};
CC KM=800 uM for glutathione (at pH 6.8) {ECO:0000269|PubMed:17124497,
CC ECO:0000269|PubMed:7775463};
CC KM=218 uM for glutathione {ECO:0000269|PubMed:17124497,
CC ECO:0000269|PubMed:7775463};
CC KM=60 uM for spermidine (at pH 6.8) {ECO:0000269|PubMed:17124497,
CC ECO:0000269|PubMed:7775463};
CC KM=20 uM for spermidine (at pH 7.5) {ECO:0000269|PubMed:17124497,
CC ECO:0000269|PubMed:7775463};
CC KM=76 uM for spermidine {ECO:0000269|PubMed:17124497,
CC ECO:0000269|PubMed:7775463};
CC KM=900 uM for glutathionylspermidine (at pH 7.5)
CC {ECO:0000269|PubMed:17124497, ECO:0000269|PubMed:7775463};
CC Note=kcat is 7 sec(-1) for Gsp synthetase activity at pH 6.8 and 2.1
CC sec(-1) for Gsp amidase activity at pH 7.5.
CC {ECO:0000269|PubMed:7775463};
CC pH dependence:
CC Optimum pH is around 6.8 for Gsp synthetase activity.
CC {ECO:0000269|PubMed:7775463};
CC -!- PATHWAY: Sulfur metabolism; glutathione metabolism.
CC -!- PATHWAY: Amine and polyamine metabolism; spermidine metabolism.
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:17124497,
CC ECO:0000269|PubMed:7775463}.
CC -!- INTERACTION:
CC P0AES0; P31574: fixB; NbExp=2; IntAct=EBI-557080, EBI-554030;
CC P0AES0; P0AG30: rho; NbExp=5; IntAct=EBI-557080, EBI-545468;
CC -!- INDUCTION: Expression level is unaffected by H(2)O(2); however Gsp
CC rapidly accumulates in E.coli in the presence of H(2)O(2).
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; liberation of the amidase
CC domain from the synthetase domain highly activates the amidase
CC activity. {ECO:0000269|PubMed:17124497, ECO:0000269|PubMed:8999955}.
CC -!- PTM: Oxidation of Cys-59 to sulfenic acid during oxidative stress
CC selectively inhibits the amidase activity which leads to a rapid
CC increase in the amounts of intracellular Gsp and Gsp S-thiolated
CC proteins (GspSSPs). {ECO:0000269|PubMed:20530482}.
CC -!- DISRUPTION PHENOTYPE: Cells lacking this gene do not produce Gsp under
CC anaerobic conditions. Cells lacking both this gene and glutaredoxin
CC (grxA or grxB) become hypersensitive to H(2)O(2); they are even more
CC susceptible to oxidative damage than the single mutant lacking
CC glutaredoxin only. {ECO:0000269|PubMed:20530482,
CC ECO:0000269|PubMed:23097746}.
CC -!- MISCELLANEOUS: Gsp forms mixed disulfides with the thiols of a variety
CC of E.coli proteins. These mixed disulfides represent a previously
CC uncharacterized type of post-translational modification. The level of
CC these proteins is increased by oxidative stress, which implies that Gsp
CC might protect protein thiols against irreversible oxidation
CC (PubMed:20530482). {ECO:0000305|PubMed:20530482}.
CC -!- MISCELLANEOUS: No metal ion is required for the amidase activity.
CC {ECO:0000305|PubMed:8999955}.
CC -!- MISCELLANEOUS: Gsp hydrolysis to GSH and spermidine proceeds with
CC formation of a glutathionyl acyl-enzyme intermediate, utilizing a
CC cysteine residue as the catalytic nucleophile (PubMed:9398217). For Gsp
CC synthesis, GSH is likely phosphorylated at one of two GSH-binding sites
CC to form an acylphosphate intermediate that then translocates to the
CC other site for subsequent nucleophilic addition of spermidine
CC (PubMed:17124497). {ECO:0000305|PubMed:17124497,
CC ECO:0000305|PubMed:9398217}.
CC -!- SIMILARITY: In the C-terminal section; belongs to the
CC glutathionylspermidine synthase preATP-grasp family. {ECO:0000305}.
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DR EMBL; U23148; AAC43339.1; -; Genomic_DNA.
DR EMBL; U28377; AAA69155.1; -; Genomic_DNA.
DR EMBL; U00096; AAC76024.1; -; Genomic_DNA.
DR EMBL; AP009048; BAE77049.1; -; Genomic_DNA.
DR PIR; A57538; A57538.
DR RefSeq; NP_417462.1; NC_000913.3.
DR RefSeq; WP_001297309.1; NZ_SSZK01000023.1.
DR PDB; 2IO7; X-ray; 2.70 A; A/B=1-619.
DR PDB; 2IO8; X-ray; 2.10 A; A/B=1-619.
DR PDB; 2IO9; X-ray; 2.20 A; A/B=1-619.
DR PDB; 2IOA; X-ray; 2.80 A; A/B=1-619.
DR PDB; 2IOB; X-ray; 2.20 A; A/B=1-619.
DR PDB; 3A2Y; X-ray; 1.95 A; A=1-197.
DR PDB; 3A2Z; X-ray; 1.50 A; A=1-197.
DR PDB; 3A30; X-ray; 2.20 A; A=1-197.
DR PDB; 3O98; X-ray; 2.80 A; A/B=1-619.
DR PDBsum; 2IO7; -.
DR PDBsum; 2IO8; -.
DR PDBsum; 2IO9; -.
DR PDBsum; 2IOA; -.
DR PDBsum; 2IOB; -.
DR PDBsum; 3A2Y; -.
DR PDBsum; 3A2Z; -.
DR PDBsum; 3A30; -.
DR PDBsum; 3O98; -.
DR AlphaFoldDB; P0AES0; -.
DR SMR; P0AES0; -.
DR BioGRID; 4261180; 18.
DR BioGRID; 851792; 4.
DR DIP; DIP-36018N; -.
DR IntAct; P0AES0; 14.
DR STRING; 511145.b2988; -.
DR MEROPS; C51.A01; -.
DR jPOST; P0AES0; -.
DR PaxDb; P0AES0; -.
DR PRIDE; P0AES0; -.
DR EnsemblBacteria; AAC76024; AAC76024; b2988.
DR EnsemblBacteria; BAE77049; BAE77049; BAE77049.
DR GeneID; 66673115; -.
DR GeneID; 947474; -.
DR KEGG; ecj:JW2956; -.
DR KEGG; eco:b2988; -.
DR PATRIC; fig|1411691.4.peg.3741; -.
DR EchoBASE; EB2720; -.
DR eggNOG; COG0754; Bacteria.
DR HOGENOM; CLU_478805_0_0_6; -.
DR InParanoid; P0AES0; -.
DR OMA; YMGYKWQ; -.
DR PhylomeDB; P0AES0; -.
DR BioCyc; EcoCyc:GSP-MON; -.
DR BioCyc; MetaCyc:GSP-MON; -.
DR BRENDA; 3.5.1.78; 2026.
DR BRENDA; 6.3.1.8; 2026.
DR UniPathway; UPA00204; -.
DR UniPathway; UPA00819; -.
DR EvolutionaryTrace; P0AES0; -.
DR PRO; PR:P0AES0; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005829; C:cytosol; IDA:EcoCyc.
DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-KW.
DR GO; GO:0008884; F:glutathionylspermidine amidase activity; IDA:EcoCyc.
DR GO; GO:0008885; F:glutathionylspermidine synthase activity; IDA:EcoCyc.
DR GO; GO:0016874; F:ligase activity; IBA:GO_Central.
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 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 Pfam; PF05257; CHAP; 1.
DR Pfam; PF03738; GSP_synth; 1.
DR SUPFAM; SSF52440; SSF52440; 1.
DR SUPFAM; SSF54001; SSF54001; 1.
DR PROSITE; PS50911; CHAP; 1.
PE 1: Evidence at protein level;
KW 3D-structure; ATP-binding; Direct protein sequencing; Hydrolase; Ligase;
KW Magnesium; Metal-binding; Multifunctional enzyme; Nucleotide-binding;
KW Oxidation; Reference proteome.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:7775463"
FT CHAIN 2..619
FT /note="Bifunctional glutathionylspermidine
FT synthetase/amidase"
FT /id="PRO_0000070443"
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:0000269|PubMed:9398217"
FT BINDING 58
FT /ligand="glutathionylspermidine"
FT /ligand_id="ChEBI:CHEBI:57835"
FT BINDING 64
FT /ligand="glutathionylspermidine"
FT /ligand_id="ChEBI:CHEBI:57835"
FT BINDING 78..81
FT /ligand="glutathionylspermidine"
FT /ligand_id="ChEBI:CHEBI:57835"
FT BINDING 149
FT /ligand="glutathionylspermidine"
FT /ligand_id="ChEBI:CHEBI:57835"
FT BINDING 316..318
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT BINDING 316
FT /ligand="glutathione"
FT /ligand_id="ChEBI:CHEBI:57925"
FT BINDING 318
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT BINDING 330
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT BINDING 330
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT BINDING 332
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT BINDING 335
FT /ligand="glutathione"
FT /ligand_id="ChEBI:CHEBI:57925"
FT BINDING 391
FT /ligand="spermidine"
FT /ligand_id="ChEBI:CHEBI:57834"
FT BINDING 392
FT /ligand="glutathione"
FT /ligand_id="ChEBI:CHEBI:57925"
FT BINDING 446
FT /ligand="glutathione"
FT /ligand_id="ChEBI:CHEBI:57925"
FT BINDING 498
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT BINDING 533
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT BINDING 539..540
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT BINDING 568..571
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT BINDING 582
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT BINDING 603..605
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT BINDING 610
FT /ligand="spermidine"
FT /ligand_id="ChEBI:CHEBI:57834"
FT SITE 131
FT /note="Increases nucleophilicity of active site Cys; for
FT amidase activity"
FT SITE 316
FT /note="Transition state stabilizer; for synthetase
FT activity"
FT MOD_RES 59
FT /note="Cysteine sulfenic acid (-SOH); transient"
FT /evidence="ECO:0000269|PubMed:20530482"
FT MUTAGEN 59
FT /note="C->A: Loss of amidase activity."
FT /evidence="ECO:0000269|PubMed:9398217"
FT MUTAGEN 173
FT /note="C->A: No effect on amidase activity."
FT /evidence="ECO:0000269|PubMed:9398217"
FT MUTAGEN 316
FT /note="R->E: Loss of synthetase activity."
FT MUTAGEN 335
FT /note="S->A: 3.6-fold decrease in GSH affinity, 1.6-fold
FT decrease in spermidine activity, and 1.3-fold decrease in
FT synthetase activity."
FT /evidence="ECO:0000269|PubMed:17124497"
FT MUTAGEN 337
FT /note="S->A: No effect on GSH and spermidine affinity, but
FT 2-fold decrease in synthetase activity."
FT /evidence="ECO:0000269|PubMed:17124497"
FT MUTAGEN 338
FT /note="C->A: 10-fold decrease in GSH affinity, 5-fold
FT decrease in spermidine activity, but no effect on
FT synthetase activity."
FT /evidence="ECO:0000269|PubMed:17124497"
FT MUTAGEN 391
FT /note="E->A: 2-fold decrease in GSH affinity, 60-fold
FT decrease in spermidine activity, and 10-fold decrease in
FT synthetase activity."
FT /evidence="ECO:0000269|PubMed:17124497"
FT MUTAGEN 392
FT /note="E->A: 33-fold decrease in GSH affinity, 13-fold
FT decrease in spermidine activity, and 6-fold decrease in
FT synthetase activity."
FT /evidence="ECO:0000269|PubMed:17124497"
FT MUTAGEN 441
FT /note="T->A: 3-fold decrease in GSH affinity, 21-fold
FT decrease in spermidine activity, and 17-fold decrease in
FT synthetase activity."
FT /evidence="ECO:0000269|PubMed:17124497"
FT MUTAGEN 538
FT /note="R->A: 6-fold decrease in GSH affinity, 2.4-fold
FT decrease in spermidine activity, and 4-fold decrease in
FT synthetase activity."
FT /evidence="ECO:0000269|PubMed:17124497"
FT MUTAGEN 598
FT /note="R->A: 10-fold increase in GSH affinity, 9-fold
FT decrease in spermidine activity, and 15-fold decrease in
FT synthetase activity."
FT /evidence="ECO:0000269|PubMed:17124497"
FT STRAND 15..19
FT /evidence="ECO:0007829|PDB:3A2Z"
FT TURN 20..22
FT /evidence="ECO:0007829|PDB:3A2Z"
FT STRAND 23..26
FT /evidence="ECO:0007829|PDB:3A2Z"
FT HELIX 35..40
FT /evidence="ECO:0007829|PDB:3A2Z"
FT HELIX 42..44
FT /evidence="ECO:0007829|PDB:3A2Z"
FT STRAND 45..48
FT /evidence="ECO:0007829|PDB:3A2Z"
FT STRAND 51..55
FT /evidence="ECO:0007829|PDB:3A2Z"
FT HELIX 59..71
FT /evidence="ECO:0007829|PDB:3A2Z"
FT STRAND 72..74
FT /evidence="ECO:0007829|PDB:3A2Z"
FT HELIX 81..86
FT /evidence="ECO:0007829|PDB:3A2Z"
FT STRAND 89..92
FT /evidence="ECO:0007829|PDB:3A2Z"
FT TURN 93..95
FT /evidence="ECO:0007829|PDB:3A2Z"
FT STRAND 98..100
FT /evidence="ECO:0007829|PDB:3A2Z"
FT STRAND 102..105
FT /evidence="ECO:0007829|PDB:3A2Z"
FT STRAND 108..110
FT /evidence="ECO:0007829|PDB:2IOB"
FT STRAND 117..120
FT /evidence="ECO:0007829|PDB:3A2Z"
FT HELIX 124..126
FT /evidence="ECO:0007829|PDB:3A2Z"
FT TURN 127..129
FT /evidence="ECO:0007829|PDB:3A2Z"
FT STRAND 131..138
FT /evidence="ECO:0007829|PDB:3A2Z"
FT STRAND 140..146
FT /evidence="ECO:0007829|PDB:3A2Z"
FT STRAND 148..150
FT /evidence="ECO:0007829|PDB:3A2Z"
FT STRAND 162..170
FT /evidence="ECO:0007829|PDB:3A2Z"
FT STRAND 173..177
FT /evidence="ECO:0007829|PDB:3A2Z"
FT STRAND 179..182
FT /evidence="ECO:0007829|PDB:3A2Z"
FT STRAND 185..192
FT /evidence="ECO:0007829|PDB:3A2Z"
FT HELIX 206..209
FT /evidence="ECO:0007829|PDB:2IO8"
FT STRAND 212..215
FT /evidence="ECO:0007829|PDB:2IO8"
FT STRAND 229..231
FT /evidence="ECO:0007829|PDB:2IOA"
FT HELIX 232..241
FT /evidence="ECO:0007829|PDB:2IO8"
FT STRAND 245..247
FT /evidence="ECO:0007829|PDB:2IO8"
FT STRAND 251..256
FT /evidence="ECO:0007829|PDB:2IO8"
FT HELIX 257..283
FT /evidence="ECO:0007829|PDB:2IO8"
FT HELIX 285..288
FT /evidence="ECO:0007829|PDB:2IO8"
FT HELIX 289..291
FT /evidence="ECO:0007829|PDB:2IO8"
FT HELIX 295..297
FT /evidence="ECO:0007829|PDB:2IO8"
FT HELIX 298..307
FT /evidence="ECO:0007829|PDB:2IO8"
FT HELIX 309..311
FT /evidence="ECO:0007829|PDB:2IOB"
FT STRAND 314..322
FT /evidence="ECO:0007829|PDB:2IO8"
FT STRAND 325..332
FT /evidence="ECO:0007829|PDB:2IO8"
FT HELIX 339..343
FT /evidence="ECO:0007829|PDB:2IO8"
FT HELIX 345..353
FT /evidence="ECO:0007829|PDB:2IO8"
FT TURN 361..364
FT /evidence="ECO:0007829|PDB:2IO8"
FT HELIX 365..374
FT /evidence="ECO:0007829|PDB:2IO8"
FT STRAND 380..386
FT /evidence="ECO:0007829|PDB:2IO8"
FT HELIX 390..405
FT /evidence="ECO:0007829|PDB:2IO8"
FT STRAND 409..416
FT /evidence="ECO:0007829|PDB:2IO8"
FT STRAND 423..425
FT /evidence="ECO:0007829|PDB:2IO8"
FT STRAND 437..442
FT /evidence="ECO:0007829|PDB:2IO8"
FT HELIX 444..453
FT /evidence="ECO:0007829|PDB:2IO8"
FT TURN 456..458
FT /evidence="ECO:0007829|PDB:2IOB"
FT STRAND 459..461
FT /evidence="ECO:0007829|PDB:2IOB"
FT HELIX 475..479
FT /evidence="ECO:0007829|PDB:2IO8"
FT STRAND 485..488
FT /evidence="ECO:0007829|PDB:2IO8"
FT HELIX 490..493
FT /evidence="ECO:0007829|PDB:2IO8"
FT TURN 494..496
FT /evidence="ECO:0007829|PDB:2IO8"
FT HELIX 500..507
FT /evidence="ECO:0007829|PDB:2IO8"
FT STRAND 517..520
FT /evidence="ECO:0007829|PDB:2IO8"
FT HELIX 523..528
FT /evidence="ECO:0007829|PDB:2IO8"
FT STRAND 530..534
FT /evidence="ECO:0007829|PDB:2IO8"
FT TURN 539..542
FT /evidence="ECO:0007829|PDB:2IO8"
FT STRAND 544..546
FT /evidence="ECO:0007829|PDB:2IO8"
FT STRAND 552..555
FT /evidence="ECO:0007829|PDB:2IO8"
FT TURN 559..562
FT /evidence="ECO:0007829|PDB:2IO8"
FT STRAND 565..569
FT /evidence="ECO:0007829|PDB:2IO8"
FT STRAND 579..588
FT /evidence="ECO:0007829|PDB:2IO8"
FT STRAND 591..604
FT /evidence="ECO:0007829|PDB:2IO8"
FT STRAND 609..612
FT /evidence="ECO:0007829|PDB:2IO8"
FT STRAND 614..617
FT /evidence="ECO:0007829|PDB:2IO8"
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
