Any feedback?
Please rate this page
(enzyme.php)
(0/150)

BRENDA support

BRENDA Home
show all | hide all No of entries

Information on EC 6.3.1.8 - glutathionylspermidine synthase and Organism(s) Crithidia fasciculata and UniProt Accession P90518

for references in articles please use BRENDA:EC6.3.1.8
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree
IUBMB Comments
Requires magnesium ions. Involved in the synthesis of trypanothione in trypanosomatids. The enzyme from Escherichia coli is bifunctional and also catalyses the glutathionylspermidine amidase (EC 3.5.1.78) reaction, resulting in a net hydrolysis of ATP.
Specify your search results
Select one or more organisms in this record: ?
This record set is specific for:
Crithidia fasciculata
UNIPROT: P90518
Show additional data
Do not include text mining results
Include (text mining) results
Include results (AMENDA + additional results, but less precise)
Word Map
The taxonomic range for the selected organisms is: Crithidia fasciculata
The enzyme appears in selected viruses and cellular organisms
Synonyms
glutathionylspermidine synthetase, gsp synthetase, ligsps, glutathionylspermidine synthase, bifunctional glutathionylspermidine synthetase/amidase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Gen Bank AE000381-derived protein GI 1789361
-
-
-
-
GenBank U66520-derived protein GI 1813514
-
-
-
-
Glutathione:spermidine ligase (ADP-forming)
-
-
-
-
Glutathione:spermidine ligase [ADP-forming]
-
-
-
-
Glutathionylspermidine synthetase (Crithidia fasciculata strain HS6 gene Cf-GSS)
-
-
-
-
Glutathionylspermidine synthetase (Crithidia fasciculata)
-
-
-
-
Glutathionylspermidine synthetase/amidase
-
-
-
-
Gsp synthetase
-
-
-
-
Protein (Escherichia coli strain K12-MG1655 gene gsp)
-
-
-
-
Synthetase, glutathionylspermidine
-
-
-
-
Synthetase, glutathionylspermidine (Crithidia fasciculata fragment)
-
-
-
-
Synthetase, glutathionylspermidine (Crithidia fasciculata strain HS6 gene Cf-GSS)
-
-
-
-
Synthetase, glutathionylspermidine (Crithidia fasciculata)
-
-
-
-
Synthetase, glutathionylspermidine (Escherichia coli clone pJBM1 gene gsp)
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
glutathione + spermidine + ATP = glutathionylspermidine + ADP + phosphate
show the reaction diagram
glutathione + spermidine + ATP = glutathionylspermidine + ADP + phosphate
show the reaction diagram
the reaction is catalyzed by trypanothione synthase, EC 6.3.1.9, and glutathionylspermidine synthase, EC 6.3.1.8, in Crithidia fasciculata
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Acid amide formation
-
-
-
-
PATHWAY SOURCE
PATHWAYS
-
-
SYSTEMATIC NAME
IUBMB Comments
gamma-L-glutamyl-L-cysteinyl-glycine:spermidine ligase (ADP-forming) [spermidine is numbered so that atom N-1 is in the amino group of the aminopropyl part of the molecule]
Requires magnesium ions. Involved in the synthesis of trypanothione in trypanosomatids. The enzyme from Escherichia coli is bifunctional and also catalyses the glutathionylspermidine amidase (EC 3.5.1.78) reaction, resulting in a net hydrolysis of ATP.
CAS REGISTRY NUMBER
COMMENTARY hide
168257-36-1
synthetase, glutathionylspermidine (Escherichia coli clone pJBM1 gene gsp), GenBank AE000381-derived protein GI 1789361, protein (Escherichia coli strain K12-MG1655 gene gsp)
190209-90-6
synthetase, glutathionylspermidine (Crithidia fasciculata), glutathionylspermidine synthetase (Crithidia fasciculata)
209273-40-5
synthetase, glutathionylspermidine (Crithidia fasciculata fragment)
213260-31-2
synthetase, glutathionylspermidine (Crithidia fasciculata strain HS6 gene Cf-GSS), genBank U66520-derived protein GI 1813514, glutathionylspermidine synthetase (Crithidia fasciculata strain HS6 gene Cf-GSS)
9077-09-2
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
gamma-L-Glu-L-Cys-Gly + spermidine + MgATP2-
N1-(gamma-L-Glu-L-Cys-Gly)-spermidine + MgADP- + phosphate
show the reaction diagram
glutathione + spermidine + ATP
glutathionylspermidine + ADP + phosphate
show the reaction diagram
glutathionylspermidine + H2O
glutathione + spermidine
show the reaction diagram
amidase activity
-
?
2 glutathione + spermidine + 2 ATP
N1,N8-bis(glutathionyl)spermidine + 2 ADP + 2 phosphate
show the reaction diagram
trypanothione synthase overall reaction
-
-
?
gamma-Glu-Gly-Gly + spermidine + ATP
N1-(gamma-Glu-Gly-Gly)-spermidine + ADP + phosphate
show the reaction diagram
-
at 14% of the activity with gamma-L-Glu-L-Cys-Gly
-
-
?
gamma-Glu-L-Cys(S-butyl)-Gly + spermidine + ATP
N1-(gamma-Glu-L-Cys(S-butyl)-Gly)-spermidine + ADP + phosphate
show the reaction diagram
-
at 24% of the activity with gamma-L-Glu-L-Cys-Gly
-
-
?
gamma-Glu-L-Cys(S-ethyl)-Gly + spermidine + ATP
N1-(gamma-Glu-L-Cys(S-ethyl)-Gly)-spermidine + ADP + phosphate
show the reaction diagram
-
at 77% of the activity with gamma-L-Glu-L-Cys-Gly
-
-
?
gamma-Glu-L-Cys(S-propyl)-Gly + spermidine + ATP
N1-(gamma-Glu-L-Cys(S-propyl)-Gly)-spermidine + ADP + phosphate
show the reaction diagram
-
at 56% of the activity with gamma-L-Glu-L-Cys-Gly
-
-
?
gamma-Glu-L-Ser-Gly + spermidine + ATP
N1-(gamma-Glu-L-Ser-Gly)-spermidine + ADP + phosphate
show the reaction diagram
-
at 16% of the activity with gamma-L-Glu-L-Cys-Gly
-
-
?
gamma-L-Glu-L-Ala-Gly + spermidine + ATP
N1-(gamma-L-Glu-L-Ala-Gly)-spermidine + ADP + phosphate
show the reaction diagram
-
at 28% of the activity with gamma-L-Glu-L-Cys-Gly
-
-
?
gamma-L-Glu-L-alpha-aminobutyryl-Gly + spermidine + ATP
N1-(gamma-L-Glu-L-alpha-aminobutyryl-Gly)-spermidine
show the reaction diagram
-
gamma-L-Glu-L-alpha-aminobutyryl-Gly i.e. ophthalmic acid
-
-
?
gamma-L-Glu-L-Cys(S-methyl)-Gly + spermidine + ATP
N1-(gamma-L-Glu-L-Cys(S-methyl)-Gly)-spermidine + ADP + phosphate
show the reaction diagram
-
at 74% of the activity with gamma-L-Glu-L-Cys-Gly
-
-
?
gamma-L-Glu-L-Cys-Gly + spermidine + ATP
N1-(gamma-L-Glu-L-Cys-Gly)-spermidine + ADP + phosphate
show the reaction diagram
gamma-L-Glu-L-Cys-Gly + spermidine + MgATP2-
N1-(gamma-L-Glu-L-Cys-Gly)-spermidine + MgADP- + phosphate
show the reaction diagram
-
substitution of the glycine part abolishes activity
-
?
gamma-L-Glu-L-Ile-Gly + spermidine + ATP
N1-(gamma-L-Glu-L-Ile-Gly)-spermidine + ADP + phosphate
show the reaction diagram
-
at 70% of the activity with gamma-L-Glu-L-Cys-Gly
-
-
?
gamma-L-Glu-L-Val-Gly + spermidine + ATP
N1-(gamma-L-Glu-L-Val-Gly)-spermidine + ADP + phosphate
show the reaction diagram
-
at 88% of the activity with gamma-L-Glu-L-Cys-Gly
-
-
?
gamma-L-Glutamyl-L-cysteinyl-glycine + spermidine + ATP
?
show the reaction diagram
glutathione + spermidine + ATP
glutathionylspermidine + ADP + phosphate
show the reaction diagram
trypanothione synthase
-
-
?
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
glutathione + spermidine + ATP
glutathionylspermidine + ADP + phosphate
show the reaction diagram
gamma-L-Glutamyl-L-cysteinyl-glycine + spermidine + ATP
?
show the reaction diagram
glutathione + spermidine + ATP
glutathionylspermidine + ADP + phosphate
show the reaction diagram
trypanothione synthase
-
-
?
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
gamma-L-glutamyl-L-cysteinyl-diaminopropionic acid
-
-
Phosphinate
behaves as a slow-binding bisubstrate inhibitor, competitive with respect to GSH and spermidine
phosphinate analogue of glutathionylspermidine
slow tight-binding inhibition
phosphonate
acts as a a simple ground state analogue of glutathione
phosphonate analogue of glutathionylspermidine
classical, linear competitive inhibition with respect to GSH
gamma-Aminobutyryl-L-Ala-Gly
-
5 mM, 10% inhibition
gamma-L-Glu-D-Phe-Gly
-
5 mM, 10% inhibition
gamma-L-Glu-L-Leu
-
5 mM, 10% inhibition
gamma-L-Glu-L-Leu-Gly
-
5 mM, 58% inhibition
gamma-L-Glu-L-leu-L-Ala
-
5 mM, 95% inhibition
gamma-L-Glu-L-Phe-Gly
-
5 mM, 73% inhibition
gamma-L-Glu-L-Ser-Gly
-
5 mM, 50% inhibition
gamma-L-Glu-L-Val
-
5 mM, 10% inhibition
gamma-L-Glu-L-Val-L-Ala
-
5 mM, 58% inhibition
gamma-L-glutamyl-L-cysteinyl-diaminopropionic acid
-
-
Glutaryl-L-Val-Gly
-
5 mM, 10% inhibition
L-beta-Asp-L-Leu-Gly
-
5 mM, 42% inhibition
L-beta-Asp-L-Val-Gly
-
5 mM, 15% inhibition
L-Cys-Gly
-
5 mM, 20% inhibition
reduced trypanothione
-
-
additional information
-
a series of glutathione analogs where the glycine moiety is substituted for other amino acids can act as inhibitors
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.215
ATP
pH 7.2, 25°C
0.171 - 0.609
glutathione
0.5
glutathionylspermidine
pH 7.3, 25°C, amidase activity
0.096 - 0.114
MgADP-
0.059 - 0.157
spermidine
0.052
ATP
pH 7.2, 25°C, recombinant trypanothione synthase performing the glutathionylspermidine synthase reaction
14
gamma-Glu-L-Cys(S-butyl)-Gly
-
-
0.18
gamma-Glu-L-Cys(S-ethyl)-Gly
-
-
0.53
gamma-Glu-L-Cys(S-propyl)-Gly
-
-
4
gamma-L-Glu-L-Ala-Gly
-
-
0.13
gamma-L-Glu-L-Cys(S-methyl)-Gly
-
-
0.91
gamma-L-Glu-L-Cys-Gly
-
-
0.4
gamma-L-Glu-L-Ile-Gly
-
-
0.67
gamma-L-Glu-L-Val-Gly
-
-
1.175
glutathione
pH 7.2, 25°C, recombinant trypanothione synthase performing the glutathionylspermidine synthase reaction
7.424
spermidine
pH 7.2, 25°C, recombinant trypanothione synthase performing the glutathionylspermidine synthase reaction
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
9.2
gamma-Glu-L-Cys(S-butyl)-Gly
-
-
11.3
gamma-Glu-L-Cys(S-ethyl)-Gly
-
-
8.7
gamma-Glu-L-Cys(S-propyl)-Gly
-
-
2.5
gamma-L-Glu-L-Ala-Gly
-
-
8.8
gamma-L-Glu-L-Cys(S-methyl)-Gly
-
-
15
gamma-L-Glu-L-Cys-Gly
-
-
10
gamma-L-Glu-L-Ile-Gly
-
-
11.8
gamma-L-Glu-L-Val-Gly
-
-
4.2
spermidine
pH 7.2, 25°C, recombinant trypanothione synthase performing the glutathionylspermidine synthase reaction
additional information
additional information
-
-
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
7.2
gamma-L-glutamyl-L-cysteinyl-diaminopropionic acid
-
-
0.029
Phosphinate
pH 7.2, 25°C
0.019
phosphinate analogue of glutathionylspermidine
pH 7.2, 25°C
0.156
phosphonate analogue of glutathionylspermidine
pH 7.2, 25°C
5
gamma-L-Glu-L-Ser-Gly
-
-
additional information
additional information
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.072
Phosphinate
Crithidia fasciculata
pH 7.2, 25°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.9
pH 7.3, 25°C, synthetase activity
2
about, purified recombinant trypanothione synthase performing the glutathionylspermidine synthase reaction
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.6 - 4.7
isoelectric focusing
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
GSP_CRIFA
719
0
80322
Swiss-Prot
other Location (Reliability: 2)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
80340
predicted from sequence
80520
MALDI-TOF MS
80640
MALDI-TOF MS
85200
gel filtration
86000
1 * 86000, SDS-PAGE
90000
1 * 90000, SDS-PAGE
90000
-
x * 90000, SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
?
-
x * 90000, SDS-PAGE
additional information
-
in vivo a complex between EC 6.3.1.8 and EC 6.3.1.9 exists that persists during purification
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C79A
mutant retains synthetase activity, but loses amidase activity
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80°C, 4 months, loses less than 10% synthetase activity
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) to over 98% purity
using Ni-chelating and anion-exchange chromatography
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression of a recombinant histidine-tagged enzyme in Escherichia coli
gene gspS, DNA and amino acid sequence determination and analysis, expression as His-tagged enzyme in Escherichia coli strain BL21(DE3)
expression in Escherichia coli and Saccharomyces cerevisiae
overexpression in Escherichia coli and Saccharomyces cerevisiae of the gene Cf-GSS yields insoluble proteins
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pharmacology
-
the enzyme would serve as a potential target for antiparasitic chemotherapy
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Nadeau, K.C.
Biochemical studies on protein folding chaperones (HSP90 and cyclophilin) and on trypanosomal enzymes (trypanothione and glutathionylspermidine synthetases) (heat shock protein)
Diss. Abstr. Int. B
56
3744
1995
Crithidia fasciculata
-
Manually annotated by BRENDA team
Smith, K.; Nadeau, K.; Bradley, M.; Walsh, C.; Fairlamb, A.H.
Purification of glutathionylspermidine and trypanothione synthetase from Crithidia fasciculata
Protein Sci.
1
874-883
1992
Crithidia fasciculata
Manually annotated by BRENDA team
Tetaud, E.; Manai, F.; Barrett, M.P.; Nadeau, K.; Walsh, C.T.; Fairlamb, A.H.
Cloning and characterization of the two enzymes responsible for trypanothione biosynthesis in Crithidia fasciculata
J. Biol. Chem.
273
19383-19390
1998
Crithidia fasciculata, Crithidia fasciculata (P90518)
Manually annotated by BRENDA team
De Craecker, S.; Verbruggen, C.; Rajan, P.; Smith, K.; Haemers, A.; Fairlamb, A.H.
Characterization of the peptide substrate specificity of glutathionylspermidine synthetase from Crithidia fasciculata
Mol. Biochem. Parasitol.
84
25-32
1997
Crithidia fasciculata
Manually annotated by BRENDA team
Oza, S.L.; Ariyanayagam, M.R.; Fairlamb, A.H.
Characterization of recombinant glutathionylspermidine synthetase/amidase from Crithidia fasciculata
Biochem. J.
364
679-686
2002
Crithidia fasciculata (P90518), Crithidia fasciculata
Manually annotated by BRENDA team
Amssoms, K.; Oza, S.L.; Augustyns, K.; Yamani, A.; Lambeir, A.M.; Bal, G.; Van der Veken, P.; Fairlamb, A.H.; Haemers, A.
Glutathione-like tripeptides as inhibitors of glutathionylspermidine synthetase. Part 2: Substitution of the glycine part
Bioorg. Med. Chem. Lett.
12
2703-2705
2002
Crithidia fasciculata
Manually annotated by BRENDA team
Comini, M.; Menge, U.; Wissing, J.; Flohe, L.
Trypanothione synthesis in Crithidia revisited
J. Biol. Chem.
280
6850-6860
2005
Crithidia fasciculata (P90518), Crithidia fasciculata (Q5DM89), Crithidia fasciculata
Manually annotated by BRENDA team
Oza, S.L.; Chen, S.; Wyllie, S.; Coward, J.K.; Fairlamb, A.H.
ATP-dependent ligases in trypanothione biosynthesis - kinetics of catalysis and inhibition by phosphinic acid pseudopeptides
FEBS J.
275
5408-5421
2008
Crithidia fasciculata (P90518), Crithidia fasciculata
Manually annotated by BRENDA team