Information on EC 6.3.1.8 - Glutathionylspermidine synthase

Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY
6.3.1.8
-
RECOMMENDED NAME
GeneOntology No.
Glutathionylspermidine synthase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
glutathione + spermidine + ATP = glutathionylspermidine + ADP + phosphate
show the reaction diagram
the reaction is catalyzed by trypanothione synthase and glutathionylspermidine synthase in Crithidia fasciculata; the reaction is catalyzed by trypanothione synthase, EC 6.3.1.9, and glutathionylspermidine synthase, EC 6.3.1.8, in Crithidia fasciculata
Q5DM89
glutathione + spermidine + ATP = glutathionylspermidine + ADP + phosphate
show the reaction diagram
the kinetic mechanism obeys a rapid equilibrium random ter-ter model
-
glutathione + spermidine + ATP = glutathionylspermidine + ADP + phosphate
show the reaction diagram
; gamma-L-glutamyl-L-cysteinyl-glycine + spermidine + ATP = N1-(gamma-L-glutamyl-L-cysteinyl-glycyl)-spermidine + ADP + phosphate
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Acid amide formation
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Glutathione metabolism
-
glutathionylspermidine biosynthesis
-
Metabolic 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.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
-
-
Glutathionylspermidine synthetase (Crithidia fasciculata strain HS6 gene Cf-GSS)
-
-
-
-
Glutathionylspermidine synthetase (Crithidia fasciculata)
-
-
-
-
Glutathionylspermidine synthetase/amidase
-
-
-
-
Gsp synthetase
-
-
-
-
GspS
AJ748279
-
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)
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
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)
907709-2
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
trypanothione synthase; a dual specificity enzyme
SwissProt
Manually annotated by BRENDA team
bifunctional glutathionylspermidine synthetase/amidase
UniProt
Manually annotated by BRENDA team
the enzyme is bifunctional and also catalyzes the glutathionylspermidine amidase reaction, EC 3.5.1.78, resulting in a net hydrolysis of ATP
-
-
Manually annotated by BRENDA team
the enzyme is bifunctional and also catalyzes the glutathionylspermidine amidase reaction, EC 3.5.1.78, resulting in a net hydrolysis of ATP; the two activities of the bifunctional enzyme reside in distinct domains
-
-
Manually annotated by BRENDA team
inactive gene GSPS; inactive gene GSPS
AJ748279
GenBank
Manually annotated by BRENDA team
bifunctional enzyme with glutathionylspermidine synthase and trypanothione synthase activity
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
physiological function
-
hypersensitivities of the GspSA/glutaredoxin null mutants to H2O2 support the idea that GspSA and Grx synergistically defend against oxidative damage
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2 glutathione + spermidine + 2 ATP
N1,N8-bis(glutathionyl)spermidine + 2 ADP + 2 phosphate
show the reaction diagram
Q5DM89
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 + ATP
N1-(gamma-L-Glu-L-Cys-Gly)-spermidine + ADP + phosphate
show the reaction diagram
-
-
-
-
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 + ATP
N1-(gamma-L-Glu-L-Cys-Gly)-spermidine + ADP + phosphate
show the reaction diagram
-
-
-
-
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 + ATP
N1-(gamma-L-Glu-L-Cys-Gly)-spermidine + ADP + phosphate
show the reaction diagram
-
-
-
-
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
-
-
-
?
gamma-L-Glu-L-Cys-Gly + spermidine + MgATP2-
N1-(gamma-L-Glu-L-Cys-Gly)-spermidine + MgADP- + phosphate
show the reaction diagram
-, P90518
-
-
?
gamma-L-Glu-L-Cys-Gly + spermidine + MgATP2-
N1-(gamma-L-Glu-L-Cys-Gly)-spermidine + MgADP2- + 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
-
synthesis of trypanothione, which is involved in maintaining intracellular thiol redox and in defense against oxidants
-
-
-
gamma-L-Glutamyl-L-cysteinyl-glycine + spermidine + ATP
?
show the reaction diagram
-
key enzyme that participates in the first of two similar steps of trypanothione biosynthesis
-
-
-
gamma-L-Glutamyl-L-cysteinyl-glycine + spermidine + ATP
?
show the reaction diagram
-
the enzyme is involved in the biosynthesis of the trypanosomatid-specific dithiol trypanothione
-
-
-
glutathione + spermidine + ATP
glutathionylspermidine + ADP + phosphate
show the reaction diagram
-, P0AES0
-
-
-
?
glutathione + spermidine + ATP
glutathionylspermidine + ADP + phosphate
show the reaction diagram
Q5DM89
-
-
-
?
glutathione + spermidine + ATP
glutathionylspermidine + ADP + phosphate
show the reaction diagram
-
-
-
-
?
glutathione + spermidine + ATP
glutathionylspermidine + ADP + phosphate
show the reaction diagram
Q5DM89
trypanothione synthase
-
-
?
glutathione + spermidine + ATP
glutathionylspermidine + ADP + phosphate
show the reaction diagram
-
trypanothione synthase catalyzes both the reaction of glutathionylspermidine synthase, EC 6.3.1.8, and of trypanothione synthase, EC 6.3.1.9, in Trypanosoma brucei
-
-
?
glutathione + spermidine + ATP
glutathionylspermidine + ADP + phosphate
show the reaction diagram
Q5DM89
specific for, the enzyme is distinct from the trypanothione synthase, which also catalyzes the reaction but converts glutathionylsperimidine further to trypanothione
-
-
?
glutathione + spermidine + ATP
glutathionylspermidine + ADP + phosphate
show the reaction diagram
-
glutathionylspermidine is an intermediate formed in the biosynthesis of trypanothione, an essential metabolite in defence against chemical and oxidative stress in the kinetoplastida
-
-
?
glutathionylspermidine + H2O
glutathione + spermidine
show the reaction diagram
-, P90518
amidase activity
-
?
additional information
?
-
-
important reaction in defense against chemical and oxidant stress, thiol redox homeostasis, ribonucleotide metabolism, and drug resistance in parasitic kinetoplastids
-
-
-
additional information
?
-
-
the enzyme is important for several functions, e.g. for detoxification of H2O2 and organic hyperoxids, and ribonucleotide reduction
-
-
-
additional information
?
-
Q5DM89
the trypanothione synthase also converts glutathionylspermidine further to trypanothione catalyzing the reaction of EC 6.3.1.9
-
-
-
additional information
?
-
-
model of ter-reactant mechanism of GspS catalysis and postulated overview
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
gamma-L-Glutamyl-L-cysteinyl-glycine + spermidine + ATP
?
show the reaction diagram
-
synthesis of trypanothione, which is involved in maintaining intracellular thiol redox and in defense against oxidants
-
-
-
gamma-L-Glutamyl-L-cysteinyl-glycine + spermidine + ATP
?
show the reaction diagram
-
key enzyme that participates in the first of two similar steps of trypanothione biosynthesis
-
-
-
gamma-L-Glutamyl-L-cysteinyl-glycine + spermidine + ATP
?
show the reaction diagram
-
the enzyme is involved in the biosynthesis of the trypanosomatid-specific dithiol trypanothione
-
-
-
glutathione + spermidine + ATP
glutathionylspermidine + ADP + phosphate
show the reaction diagram
Q5DM89
-, trypanothione synthase
-
-
?
glutathione + spermidine + ATP
glutathionylspermidine + ADP + phosphate
show the reaction diagram
-
trypanothione synthase catalyzes both the reaction of glutathionylspermidine synthase, EC 6.3.1.8, and of trypanothione synthase, EC 6.3.1.9, in Trypanosoma brucei
-
-
?
glutathione + spermidine + ATP
glutathionylspermidine + ADP + phosphate
show the reaction diagram
-
glutathionylspermidine is an intermediate formed in the biosynthesis of trypanothione, an essential metabolite in defence against chemical and oxidative stress in the kinetoplastida
-
-
?
additional information
?
-
-
important reaction in defense against chemical and oxidant stress, thiol redox homeostasis, ribonucleotide metabolism, and drug resistance in parasitic kinetoplastids
-
-
-
additional information
?
-
-
the enzyme is important for several functions, e.g. for detoxification of H2O2 and organic hyperoxids, and ribonucleotide reduction
-
-
-
additional information
?
-
Q5DM89
the trypanothione synthase also converts glutathionylspermidine further to trypanothione catalyzing the reaction of EC 6.3.1.9
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
ATP
Q5DM89
as MgATP2-; as MgATP2-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Mg2+
Q5DM89
as MgATP2-; as MgATP2-
Mg2+
P0AES0
two Mg ions per molecule
Mg2+
-
required
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
(2-[[([2-[(4-amino-4-carboxybutanoyl)amino]propanoyl]amino)methyl](hydroxy)phosphoryl]ethyl)(hydroxy)dioxophosphate(1-)
P0AES0
inhibits the reaction in the presence of spermidine
4-[(14-ammonio-5-hydroxy-1-methyl-5-oxido-2-oxo-6-oxa-3,10-diaza-5-phosphatetradec-1-yl)amino]-1-carboxy-4-oxobutan-1-aminium
P0AES0
-
4-[[2-([[[4-[(4-ammoniobutyl)amino]butyl](hydroxy)phosphoryl]methyl]amino)-1-methyl-2-oxoethyl]amino]-1-carboxy-4-oxobutan-1-aminium
P0AES0
-
buthionine sulfoximine
-
-
gamma-Aminobutyryl-L-Ala-Gly
-
5 mM, 10% inhibition
gamma-glutamyl-N-[[(3-aminopropoxy)(2-phosphonoethyl)phosphoryl]methyl]alaninamide
P0AES0
-
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
Hydrogen O-[3-[N-(4-Aminobutyl)amino]propyl][[(gamma-glutamylalanyl)amino]methyl]phosphonate
-
-
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
N1-glutathionylspermidine analogs
-
selective inhibitors against EC 6.3.1.8 and EC 3.5.1.78 activities provide evidence of interdomain communication in the bifunctional enzyme
Non-polyamine-containing phosphoamidate
-
-
-
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
Polyamine-containing phosphapeptides
-
potent and selective
-
reduced trypanothione
-
-
Melarsen oxide
-
-
additional information
-
a series of glutathione analogs where the glycine moiety is substituted for other amino acids can act as inhibitors
-
additional information
-
the enzyme mediates resistance to H2O2 and organic hyperoxids, and to trypanocidic drugs
-
additional information
-
H2O2 leads to inhibition of the amidase activity, while the glutathionylspermidine synthase activity is almost unaffected
-
additional information
-
design, synthesis, and biochemical evaluation of phosphonate and phosphonamidate analogs of glutathionylspermidine as inhibitors
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.052
-
ATP
Q5DM89
pH 7.2, 25C, recombinant trypanothione synthase performing the glutathionylspermidine synthase reaction
0.215
-
ATP
-
pH 7.2, 25C
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
-
-
0.0224
-
glutathione
P0AES0
mutant enzyme R598A
0.171
-
glutathione
-
pH 7.3, 25C, synthetase activity of the C79A mutant
0.218
-
glutathione
P0AES0
wild type enzyme
0.224
-
glutathione
P0AES0
mutant enzyme S337A
0.242
-
glutathione
-
pH 7.3, 25C, synthetase activity
0.251
-
glutathione
P0AES0
mutant enzyme K607A
0.407
-
glutathione
P0AES0
mutant enzyme E391A
0.609
-
glutathione
-
pH 7.2, 25C
0.691
-
glutathione
P0AES0
mutant enzyme T441A
0.794
-
glutathione
P0AES0
mutant enzyme S335A
1.175
-
glutathione
Q5DM89
pH 7.2, 25C, recombinant trypanothione synthase performing the glutathionylspermidine synthase reaction
1.324
-
glutathione
P0AES0
mutant enzyme R538A
2.098
-
glutathione
P0AES0
mutant enzyme C338A
7.298
-
glutathione
P0AES0
mutant enzyme E392A
0.5
-
glutathionylspermidine
-
pH 7.3, 25C, amidase activity
0.096
-
MgADP2-
-
pH 7.3, 25C, synthetase activity of the C79A mutant
-
0.114
-
MgADP2-
-
pH 7.3, 25C, synthetase activity
-
0.059
-
spermidine
-
pH 7.3, 25C, synthetase activity
0.076
-
spermidine
P0AES0
mutant enzyme S337A; wild type enzyme
0.078
-
spermidine
-
pH 7.3, 25C, synthetase activity of the C79A mutant
0.126
-
spermidine
P0AES0
mutant enzyme S335A
0.153
-
spermidine
P0AES0
mutant enzyme K607A
0.157
-
spermidine
-
pH 7.2, 25C
0.182
-
spermidine
P0AES0
mutant enzyme R538A
0.406
-
spermidine
P0AES0
mutant enzyme C338A
0.665
-
spermidine
P0AES0
mutant enzyme R598A
1.008
-
spermidine
P0AES0
mutant enzyme E392A
1.61
-
spermidine
P0AES0
mutant enzyme T441A
4.522
-
spermidine
P0AES0
mutant enzyme E391A
7.424
-
spermidine
Q5DM89
pH 7.2, 25C, recombinant trypanothione synthase performing the glutathionylspermidine synthase reaction
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
-
-
4.2
-
spermidine
Q5DM89
pH 7.2, 25C, recombinant trypanothione synthase performing the glutathionylspermidine synthase reaction
11.8
-
gamma-L-Glu-L-Val-Gly
-
-
additional information
-
additional information
-
-
-
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.006
-
4-[(14-ammonio-5-hydroxy-1-methyl-5-oxido-2-oxo-6-oxa-3,10-diaza-5-phosphatetradec-1-yl)amino]-1-carboxy-4-oxobutan-1-aminium
P0AES0
-
0.0000078
-
4-[[2-([[[4-[(4-ammoniobutyl)amino]butyl](hydroxy)phosphoryl]methyl]amino)-1-methyl-2-oxoethyl]amino]-1-carboxy-4-oxobutan-1-aminium
P0AES0
-
5
-
gamma-L-Glu-L-Ser-Gly
-
-
0.029
-
Phosphinate
-
pH 7.2, 25C
0.019
-
phosphinate analogue of glutathionylspermidine
-
pH 7.2, 25C
0.156
-
phosphonate analogue of glutathionylspermidine
-
pH 7.2, 25C
7.2
-
gamma-L-glutamyl-L-cysteinyl-diaminopropionic acid
-
-
additional information
-
additional information
-
inhibition kinetics and mechanism, model of ter-reactant mechanism of GspS catalysis and postulated, overview
-
IC50 VALUE [mM]
IC50 VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.072
-
Phosphinate
-
pH 7.2, 25C
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
2
-
Q5DM89
about, purified recombinant trypanothione synthase performing the glutathionylspermidine synthase reaction
7.4
-
-
-
7.9
-
-
pH 7.3, 25C, synthetase activity
additional information
-
-
-
additional information
-
-
enzyme cloned in Escherichia coli and Saccharomyces cerevisiae is inactive
additional information
-
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.2
-
Q5DM89
assay at; assay at
7.2
-
-
assay at
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
-
Q5DM89
assay at; assay at
25
-
-
assay at
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4.6
4.7
-
isoelectric focusing
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
80340
-
-
predicted from sequence
80520
-
-
MALDI-TOF MS
80640
-
-
MALDI-TOF MS
85200
-
-
gel filtration
138000
-
P0AES0
analytical ultracentrifugation
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 90000, SDS-PAGE
dimer
P0AES0
2 * 70000, analytical ultracentrifugation
monomer
-
1 * 86000, SDS-PAGE
monomer
-
1 * 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
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
hanging drop vapour diffusion method generating crystals of GspS in 0.1M TrisHCl (pH 8.5) containing 12% (v/v) PEG3350 and 0.5 M MgCl2
P0AES0
mutant C59A lacking amidase activity, in complex with amidase substrate gluthionylspermidine and ADP. Homodimer, and each monomer contains the N-terminal amidase and C-terminal synthetase domains connected by a linker in between
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-80C, 4 months, loses less than 10% synthetase activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) to over 98% purity
-
using Ni-chelating and anion-exchange chromatography
-
nickel affinity column chromatography
P0AES0
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression in Escherichia coli and Saccharomyces cerevisiae; overexpression in Escherichia coli and Saccharomyces cerevisiae of the gene Cf-GSS yields insoluble proteins
-
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)
-
expressed in Escherichia coli BL21 (DE3) cells
P0AES0
gene GSPS, a pseudogene with 2 frame shift mutations and 2 stop codons, inactive enzyme, phylogenetic analysis
AJ748279
DNA sequence determination, cloning and RNAi transfection, expression in Escherichia coli
-
enzyme expression analysis, cloning and transfection of RNAi
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
C79A
-
mutant retains synthetase activity, but loses amidase activity
C338A
P0AES0
increased Km compared to the wild type enzyme
E391A
P0AES0
increased Km compared to the wild type enzyme
E392A
P0AES0
increased Km compared to the wild type enzyme
K607A
P0AES0
increased Km compared to the wild type enzyme
R316E
P0AES0
no activity
R538A
P0AES0
increased Km compared to the wild type enzyme
R598A
P0AES0
decreased Km compared to the wild type enzyme
S335A
P0AES0
increased Km compared to the wild type enzyme
S337A
P0AES0
increased Km compared to the wild type enzyme
T441A
P0AES0
increased Km compared to the wild type enzyme
additional information
-
enzyme downregulation leads to decreased growth rate of the parasite, increased H2O2 and tert-butylhydroperoxide sensitivity, enzyme knockdown leads to depletion of both trypanothione and glutathionylspermidine, and glutathione and polyamine accumulation, RNA interference method, overview
additional information
-
enzyme downregulation leads to increased trypanocidic drug sensitivity, enzyme knockdown leads to depletion of both trypanothione and glutathionylspermidine and glutathione accumulation, RNA interference method, overview
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
pharmacology
-
the enzyme would serve as a potential target for antiparasitic chemotherapy
drug development
-
the enzyme is a target for drug development