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 27-MAR-2024, entry version 133. 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; Pseudomonadota; 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}. 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; 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; 511145-b2988; -. DR EnsemblBacteria; AAC76024; AAC76024; b2988. DR GeneID; 75205173; -. 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; WPRIRHS; -. DR OrthoDB; 9765517at2; -. DR PhylomeDB; P0AES0; -. DR BioCyc; EcoCyc:GSP-MONOMER; -. DR BioCyc; MetaCyc:GSP-MONOMER; -. 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: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 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 //