Information on EC 6.3.1.9 - trypanothione synthase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
6.3.1.9
-
RECOMMENDED NAME
GeneOntology No.
trypanothione synthase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
glutathione + glutathionylspermidine + ATP = N1,N8-bis(glutathionyl)spermidine + ADP + phosphate
show the reaction diagram
glutathione + spermidine + ATP = glutathionylspermidine + ADP + phosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Acid amide formation
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Glutathione metabolism
-
-
glutathionylspermidine biosynthesis
-
-
Metabolic pathways
-
-
trypanothione biosynthesis
-
-
glutathione metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
spermidine/glutathionylspermidine:glutathione ligase (ADP-forming)
The enzyme, characterized from several trypanosomatids (e.g. Trypanosoma cruzi) catalyses two subsequent reactions, leading to production of trypanothione from glutathione and spermidine. Some trypanosomatids (e.g. Crithidia species and some Leishmania species) also contain an enzyme that only carries out the first reaction (cf. EC 6.3.1.8, glutathionylspermidine synthase). The enzyme is bifunctional, and also catalyses the hydrolysis of glutathionylspermidine and trypanothione (cf. EC 3.5.1.78, glutathionylspermidine amidase).
CAS REGISTRY NUMBER
COMMENTARY hide
130246-69-4
-
213260-30-1
synthetase, trypanothione (Crithidia fasciculata strain HS6 gene Cf-TS) /GenBank AF006615-derived protein GI 3004644 /trypanothione synthetase (Crithidia fasciculata strain HS6 gene Cf-TS)
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
-
trypanothione synthetase catalyses the two-step biosynthesis of trypanothione from spermidine and glutathione
physiological function
-
culture of a tetracyclinedependent conditional double knockout cDKO parasites without tetracycline induction results in loss of trypanothione and accumulation of glutathione, followed by growth inhibition and cell lysis after 6 days. In the absence of inducer, cDKO cells are unable to infect mice, confirming that the enzyme is essential for virulence in vivo as well as in vitro
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
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
gamma-L-Glu-L-alpha-aminobutyrylglycine + N1-(gamma-Glu-L-Cys-Gly)-spermidine + ATP
?
show the reaction diagram
-
gamma-L-Glu-L-alpha-aminobutyrylglycine i.e. ophthalmic acid
-
-
-
gamma-L-Glu-L-Cys-Gly + N1-(gamma-Glu-L-Cys-Gly)-spermidine + ATP
N1,N8-bis-(gamma-L-glutamyl-L-cysteinyl-glycyl)-spermidine + ADP + phosphate
show the reaction diagram
gamma-L-Glu-L-Cys-Gly + N1-(gamma-Glu-L-Cys-Gly)-spermidine + ATP + H2O
N1,N8-bis-(gamma-L-Glu-L-Cys-Gly)-spermidine + ADP + phosphate + NH3
show the reaction diagram
gamma-L-glutamyl-L-cysteinyl-glycine + N1-(gamma-L-glutamyl-L-cysteinyl-glycyl)-spermidine + ATP
N1,N8-bis-(gamma-L-glutamyl-L-cysteinyl-glycyl)-spermidine + ADP + phosphate
show the reaction diagram
glutathione + glutathionylspermidine + ATP
N1,N8-bis(glutathionyl)spermidine + ADP + phosphate
show the reaction diagram
glutathione + N8-acetylspermidine + ATP
N1-glutathionyl-N8-acetylspermidine + ADP + phosphate
show the reaction diagram
-
-
-
?
glutathione + spermidine + 2 ATP
N1,N8-bis(glutathionyl)spermidine + 2 ADP + 2 phosphate
show the reaction diagram
glutathione + spermidine + ATP
glutathionylspermidine + ADP + phosphate
show the reaction diagram
glutathione + spermidine + ATP
N1-glutathionylspermidine + N8-glutathionylspermidine + ADP + phosphate
show the reaction diagram
N1-glutathionylspermidine + glutathione + ATP
N1,N8-bis(glutathionyl)spermidine + ADP + phosphate
show the reaction diagram
N8-glutathionylspermidine + glutathione + ATP
N1,N8-bis(glutathionyl)spermidine + ADP + phosphate
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
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
gamma-L-Glu-L-Cys-Gly + N1-(gamma-Glu-L-Cys-Gly)-spermidine + ATP
N1,N8-bis-(gamma-L-glutamyl-L-cysteinyl-glycyl)-spermidine + ADP + phosphate
show the reaction diagram
gamma-L-glutamyl-L-cysteinyl-glycine + N1-(gamma-L-glutamyl-L-cysteinyl-glycyl)-spermidine + ATP
N1,N8-bis-(gamma-L-glutamyl-L-cysteinyl-glycyl)-spermidine + ADP + phosphate
show the reaction diagram
glutathione + glutathionylspermidine + ATP
N1,N8-bis(glutathionyl)spermidine + ADP + phosphate
show the reaction diagram
glutathione + spermidine + 2 ATP
N1,N8-bis(glutathionyl)spermidine + 2 ADP + 2 phosphate
show the reaction diagram
glutathione + spermidine + ATP
glutathionylspermidine + ADP + phosphate
show the reaction diagram
glutathione + spermidine + ATP
N1-glutathionylspermidine + N8-glutathionylspermidine + ADP + phosphate
show the reaction diagram
N1-glutathionylspermidine + glutathione + ATP
N1,N8-bis(glutathionyl)spermidine + ADP + phosphate
show the reaction diagram
N8-glutathionylspermidine + glutathione + ATP
N1,N8-bis(glutathionyl)spermidine + ADP + phosphate
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
(NH4)2SO4
optimal up to 50 mM
KCl
optimal up to 50 mM
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,3,5-trimethyl-N-[(5-methyl-2-phenyl-2H-1,2,3-triazol-4-yl)methyl]-1H-pyrazole-4-sulfonamide
-
-
1-(3-(1-isobutyl-1H-pyrazol-4-yl)-1H-indazol-1-yl)-3,3-dimethylbutan-2-one
-
-
1-(3-(1H-indol-4-yl)-1H-indazol-1-yl)-3,3-dimethylbutan-2-one
-
-
1-(3-(3-chlorophenyl)-1H-indazol-1-yl)-3,3-dimethylbutan-2-one
-
-
1-(3-(3-fluorophenyl)-1H-indazol-1-yl)-3,3-dimethylbutan-2-one
-
-
1-(3-(3-pyridyl)-1H-indazol-1-yl)-3,3-dimethylbutan-2-one
-
-
1-(3-(4-chlorophenyl)-1H-indazol-1-yl)-3,3-dimethylbutan-2-one
-
-
1-(3-(furan-2-yl)-1H-indazol-1-yl)-3,3-dimethylbutan-2-one
-
-
1-(3-phenyl-1H-indazol-1-yl)-3,3-dimethylbutan-2-one
-
-
1-(5-(3,5-dichlorophenyl)-2H-tetrazol-2-yl)-3,3-dimethylbutan-2-one
-
-
1-(5-(3,5-dichlorophenyl)-2H-tetrazol-2-yl)butan-2-one
-
-
1-(5-(3,5-dichlorophenyl)-2H-tetrazol-2-yl)propan-2-one
-
-
1-(5-(3,5-difluorophenyl)-2H-tetrazol-2-yl)-3,3-dimethylbutan-2-one
-
-
1-(5-(3-fluorophenyl)-2H-tetrazol-2-yl)-3,3-dimethylbutan-2-one
-
-
1-(tert-butylsulfonylmethyl)-3-(3-fluorophenyl)-1H-indazole
-
-
1-[3-(3-fluorophenyl)-1H-indazol-1-yl]-3,3-dimethylbutan-2-one
-
i.e. DDD85811, inhibits growth of bloodstream Trypanosoma brucei in vitro
1-[5-(3-chloro-4-methylphenyl)-2H-tetrazol-2-yl]-3,3-dimethylbutan-2-one
-
-
2,4-dimethyl-N-[(5-methyl-2-phenyl-2H-1,2,3-triazol-4-yl)methyl]-1,3-thiazole-5-sulfonamide
-
-
2,4-dimethyl-N-[(5-methyl-2-phenyl-2H-1,2,3-triazol-4-yl)methyl]thiophene-3-sulfonamide
-
-
2,6-dichloro-N-[(2-phenyl-1,3-thiazol-4-yl)methyl]benzenesulfonamide
-
-
2,6-dichloro-N-[(3-phenyl-1,2,4-oxadiazol-5-yl)methyl]benzenesulfonamide
-
-
2,6-difluoro-N-[(2-phenyl-1,3-thiazol-4-yl)methyl]benzenesulfonamide
-
-
2,6-dimethoxy-N-[(5-methyl-2-phenyl-2H-1,2,3-triazol-4-yl)methyl]benzenesulfonamide
-
-
2-((tert-butylsulfonyl)methyl)-4-(2-fluorophenyl)thiazole
-
-
2-((tert-butylsulfonyl)methyl)-4-(2-methylphenyl)thiazole
-
-
2-((tert-butylsulfonyl)methyl)-4-(3-chlorophenyl)thiazole
-
-
2-((tert-butylsulfonyl)methyl)-4-(3-methoxyphenyl)thiazole
-
-
2-((tert-butylsulfonyl)methyl)-4-(phenyl)thiazole
-
-
2-(3-(3-fluorophenyl)-1H-indazol-1-yl)-1-(piperidin-1-yl)ethanone
-
-
2-(3-(3-fluorophenyl)-1H-indazol-1-yl)-N,N-dimethylacetamide
-
-
2-(3-(3-fluorophenyl)-1H-indazol-1-yl)-N-(2-(4-methylpiperazin-1-yl)ethyl)acetamide
-
-
2-(3-(3-fluorophenyl)-1H-indazol-1-yl)-N-(3-(4-methylpiperazin-1-yl)propyl)acetamide
-
-
2-(5-(3,5-dichlorophenyl)-2H-tetrazol-2-yl)-N,N-dimethylacetamide
-
-
2-(5-(3,5-difluorophenyl)-2H-tetrazol-2-yl)-1-phenylethanone
-
-
2-(tert-butylsulfonylmethyl)-4-(3-trifluoromethylphenyl)thiazole
-
-
2-methyl-N-((5-methyl-2-phenyl-2H-1,2,3-triazol-4-yl)methyl)propane-2-sulfonamide
-
-
2-[3-(3-fluorophenyl)-1H-indazol-1-yl]-1-(piperidin-1-yl)ethanone
-
i.e. DDD86243, inhibits growth of bloodstream Trypanosoma brucei in vitro. Exposure to DDD86243 at 2 x EC50 for 72 h decreases intracellular trypanothione levels to less than 10% of wild type, with a corresponding 5fold increase in the precursor metabolite, glutathione
3,3-dimethyl-1-(3-(1-methyl-1H-indol-4-yl)-1H-indazol-1-yl)butan-2-one
-
-
3,3-dimethyl-1-(3-(naphthalen-2-yl)-1H-indazol-1-yl)butan-2-one
-
-
3,3-dimethyl-1-(3-(pyridin-4-yl)-1H-indazol-1-yl)butan-2-one
-
-
3,3-dimethyl-1-(3-(thiophen-3-yl)-1H-indazol-1-yl)butan-2-one
-
-
3,3-dimethyl-1-(5-phenyl-2H-tetrazol-2-yl)butan-2-one
-
-
3,3-dimethyl-1-[5-[3-(trifluoromethyl)phenyl]-2H-tetrazol-2-yl]butan-2-one
-
-
3,3-dimethyl-1-[5-[4-(trifluoromethyl)phenyl]-2H-tetrazol-2-yl]butan-2-one
-
-
3,5-dichloro-N-[(2-phenyl-1,3-thiazol-4-yl)methyl]benzenesulfonamide
-
-
3-((3-(3-fluorophenyl)-1H-indazol-1-yl)methyl)-5-methylisoxazole
-
-
3-((3-(3-fluorophenyl)-1H-indazol-1-yl)methyl)-5-phenylisoxazole
-
-
3-(3-fluorophenyl)-1-(pyridin-2-ylmethyl)-1H-indazole
-
-
3-(3-fluorophenyl)-1-(pyridin-3-ylmethyl)-1H-indazole
-
-
3-(tert-butylsulfonylmethyl)-1-(3-fluorophenyl)-1H-indazole
-
-
3-(tert-butylsulfonylmethyl)-1-phenyl-1H-indazole
-
-
4-((3-(3-fluorophenyl)-1H-indazol-1-yl)methyl)-2,5-dimethyloxazole
-
-
4-(2-hydroxyethyl)-1-piperazinepropanesulfonic acid
above 100 mM
4-acetyl-N-[(2-phenyl-1,3-thiazol-4-yl)methyl]benzenesulfonamide
-
-
4-methoxy-N-[(3-phenyl-1,2,4-oxadiazol-5-yl)methyl]benzenesulfonamide
-
-
4-methoxy-N-[[3-(2-methylphenyl)-1,2,4-oxadiazol-5-yl]methyl]benzenesulfonamide
-
-
4-methyl-N-[(2-phenyl-1,3-thiazol-4-yl)methyl]benzenesulfonamide
-
-
ammonium sulfate
almost complete inhibition at 500 mM
betulin
competitive inhibition
buthionine sulfoximine
-
-
conessine
competitive allosteric inhibition
glutathione
KCl
approx. 50% inhibition at 500 mM
Melarsen oxide
-
-
methyl (3-phenyl-1H-indazol-1-yl)acetate
-
-
N,N-dibenzyl-2-[3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl]acetamide
-
-
N,N-diethyl-2-(3-(3-fluorophenyl)-1H-indazol-1-yl)acetamide
-
-
N,N-diethyl-2-(3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl)acetamide
-
-
N-(1-phenylethyl)-2-[3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl]acetamide
-
-
N-(2-(dimethylamino)ethyl)-2-(3-(3-fluorophenyl)-1H-indazol-1-yl)acetamide
-
-
N-(2-methoxyethyl)-2-[3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl]acetamide
-
-
N-(2-phenylethyl)-2-[3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl]acetamide
-
-
N-(3-(1H-imidazol-1-yl)propyl)-2-(3-(3-fluorophenyl)-1H-indazol-1-yl)acetamide
-
-
N-(3-(dimethylamino)propyl)-2-(3-(3-fluorophenyl)-1H-indazol-1-yl)acetamide
-
-
N-(4-chlorobenzyl)-2-(3-(3-fluorophenyl)-1H-indazol-1-yl)acetamide
-
-
N-(4-chlorobenzyl)-2-(3-(trifluoromethyl)-4,5,6,7-tetrahydro-1Hindazol-1-yl)acetamide
-
-
N-(4-[methyl[(3-phenyl-1,2,4-oxadiazol-5-yl)methyl]sulfamoyl]phenyl)acetamide
-
-
N-(4-[[(3-phenyl-1,2,4-oxadiazol-5-yl)methyl]sulfamoyl]phenyl)acetamide
-
-
N-(butan-2-yl)-2-[3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl]acetamide
-
-
N-(thiophen-2-ylmethyl)-2-[3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl]acetamide
-
-
N-cyclohexyl-2-[3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl]acetamide
-
-
N-methyl-(3-(dimethylamino)propyl)-2-(3-(3-fluorophenyl)-1H-indazol-1-yl)acetamide
-
-
N-phenyl-2-[3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl]acetamide
-
-
N-propyl-2-[3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl]acetamide
-
-
N-[(2-phenyl-1,3-thiazol-4-yl)methyl]naphthalene-2-sulfonamide
-
-
N-[(3-phenyl-1,2,4-oxadiazol-5-yl)methyl]benzenesulfonamide
-
-
N-[(5-methyl-2-phenyl-2H-1,2,3-triazol-4-yl)methyl]naphthalene-1-sulfonamide
-
-
N-[(5-methyl-2-phenyl-2H-1,2,3-triazol-4-yl)methyl]thiophene-2-sulfonamide
-
-
N-[4-([[3-(2-chlorophenyl)-1,2,4-oxadiazol-5-yl]methyl]sulfamoyl)phenyl]acetamide
-
-
N-[4-([[3-(2-methylphenyl)-1,2,4-oxadiazol-5-yl]methyl]sulfamoyl)phenyl]acetamide
-
-
N-[[3-(2-chlorophenyl)-1,2,4-oxadiazol-5-yl]methyl]benzenesulfonamide
-
-
N-[[3-(2-methylphenyl)-1,2,4-oxadiazol-5-yl]methyl]benzenesulfonamide
-
-
nifurtimox
-
-
Pentamidine
-
-
prochlorperazine
-
inhibitor DDD66604, binding mode and structure, overview
tert-butyl-4-(2-(2-(3-(3-fluorophenyl)-1H-indazol-1-yl)acetamido)ethyl)piperazine-1-carboxylate
-
-
tomatine
competitive inhibition versus spermidine, uncompetitive inhibition with respect to ATP and GSH
triostam
-
-
trypanothione
-
competitive feedback inhibition
uvaol
competitive allosteric inhibition
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0066 - 0.053
ATP
0.914
gamma-L-Glu-L-Cys-Gly
-
-
0.032 - 0.407
glutathione
0.04 - 0.48
glutathionylspermidine
0.19
GSH
pH 8.0, with N1-(gamma-L-glutamyl-L-cysteinyl-glycyl)-spermidine
0.063 - 0.4
MgATP2-
0.066
N1-(gamma-L-glutamyl-L-cysteinyl-glycyl)-spermidine
pH 8.0
0.012 - 0.032
N1-glutathionylspermidine
0.007 - 0.032
N8-glutathionylspermidine
0.092 - 1.07
spermidine
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.3 - 5.2
glutathione
8.7
glutathionylspermidine
Crithidia fasciculata
Q5DM89
pH 7.2, 25C, recombinant trypanothione synthase
2.8
N1-(gamma-L-glutamyl-L-cysteinyl-glycyl)-spermidine
Trypanosoma cruzi
Q9GT49
pH 8.0
10
N1-glutathionylspermidine
Crithidia fasciculata
-
-
24.3
N8-glutathionylspermidine
Crithidia fasciculata
-
-
28.6
spermidine
Crithidia fasciculata
-
-
additional information
additional information
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0063
betulin
pH 8.0, 25C, recombinant enzyme
0.0031
conessine
pH 8.0, 25C, recombinant enzyme
0.132 - 1.085
glutathione
0.0068 - 0.0125
tomatine
0.223 - 0.377
trypanothione
0.0036
uvaol
pH 8.0, 25C, recombinant enzyme
additional information
additional information
inhibition kinetics, overview
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0026
1,3,5-trimethyl-N-[(5-methyl-2-phenyl-2H-1,2,3-triazol-4-yl)methyl]-1H-pyrazole-4-sulfonamide
Trypanosoma brucei
-
above, pH and temperature not specified in the publication
0.1
1-(3-(1-isobutyl-1H-pyrazol-4-yl)-1H-indazol-1-yl)-3,3-dimethylbutan-2-one
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0037
1-(3-(1H-indol-4-yl)-1H-indazol-1-yl)-3,3-dimethylbutan-2-one
Trypanosoma brucei
-
above, pH and temperature not specified in the publication
0.00035
1-(3-(3-chlorophenyl)-1H-indazol-1-yl)-3,3-dimethylbutan-2-one
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.00009
1-(3-(3-fluorophenyl)-1H-indazol-1-yl)-3,3-dimethylbutan-2-one
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.003
1-(3-(3-pyridyl)-1H-indazol-1-yl)-3,3-dimethylbutan-2-one
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.1
1-(3-(4-chlorophenyl)-1H-indazol-1-yl)-3,3-dimethylbutan-2-one
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0019
1-(3-(furan-2-yl)-1H-indazol-1-yl)-3,3-dimethylbutan-2-one
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.00015
1-(3-phenyl-1H-indazol-1-yl)-3,3-dimethylbutan-2-one
Trypanosoma brucei
-
above, pH and temperature not specified in the publication
0.0003
1-(5-(3,5-dichlorophenyl)-2H-tetrazol-2-yl)-3,3-dimethylbutan-2-one
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0027
1-(5-(3,5-dichlorophenyl)-2H-tetrazol-2-yl)butan-2-one
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0065
1-(5-(3,5-dichlorophenyl)-2H-tetrazol-2-yl)propan-2-one
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0019
1-(5-(3,5-difluorophenyl)-2H-tetrazol-2-yl)-3,3-dimethylbutan-2-one
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0015
1-(5-(3-fluorophenyl)-2H-tetrazol-2-yl)-3,3-dimethylbutan-2-one
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.00012
1-(tert-butylsulfonylmethyl)-3-(3-fluorophenyl)-1H-indazole
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.000095
1-[3-(3-fluorophenyl)-1H-indazol-1-yl]-3,3-dimethylbutan-2-one
Trypanosoma brucei
-
pH 8.0, 22C
0.0072
1-[5-(3-chloro-4-methylphenyl)-2H-tetrazol-2-yl]-3,3-dimethylbutan-2-one
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0022
2,4-dimethyl-N-[(5-methyl-2-phenyl-2H-1,2,3-triazol-4-yl)methyl]-1,3-thiazole-5-sulfonamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.024
2,4-dimethyl-N-[(5-methyl-2-phenyl-2H-1,2,3-triazol-4-yl)methyl]thiophene-3-sulfonamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0063
2,6-dichloro-N-[(2-phenyl-1,3-thiazol-4-yl)methyl]benzenesulfonamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.016
2,6-dichloro-N-[(3-phenyl-1,2,4-oxadiazol-5-yl)methyl]benzenesulfonamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0061
2,6-difluoro-N-[(2-phenyl-1,3-thiazol-4-yl)methyl]benzenesulfonamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.1
2,6-dimethoxy-N-[(5-methyl-2-phenyl-2H-1,2,3-triazol-4-yl)methyl]benzenesulfonamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0011
2-((tert-butylsulfonyl)methyl)-4-(2-fluorophenyl)thiazole
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0032
2-((tert-butylsulfonyl)methyl)-4-(2-methylphenyl)thiazole
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0004
2-((tert-butylsulfonyl)methyl)-4-(3-chlorophenyl)thiazole
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.012
2-((tert-butylsulfonyl)methyl)-4-(3-methoxyphenyl)thiazole
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0012
2-((tert-butylsulfonyl)methyl)-4-(phenyl)thiazole
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.00014
2-(3-(3-fluorophenyl)-1H-indazol-1-yl)-1-(piperidin-1-yl)ethanone
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.00011
2-(3-(3-fluorophenyl)-1H-indazol-1-yl)-N,N-dimethylacetamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.00086
2-(3-(3-fluorophenyl)-1H-indazol-1-yl)-N-(2-(4-methylpiperazin-1-yl)ethyl)acetamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.00083
2-(3-(3-fluorophenyl)-1H-indazol-1-yl)-N-(3-(4-methylpiperazin-1-yl)propyl)acetamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0006
2-(5-(3,5-dichlorophenyl)-2H-tetrazol-2-yl)-N,N-dimethylacetamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.045
2-(5-(3,5-difluorophenyl)-2H-tetrazol-2-yl)-1-phenylethanone
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0011
2-(tert-butylsulfonylmethyl)-4-(3-trifluoromethylphenyl)thiazole
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.037
2-methyl-N-((5-methyl-2-phenyl-2H-1,2,3-triazol-4-yl)methyl)propane-2-sulfonamide
Trypanosoma brucei
-
above, pH and temperature not specified in the publication
0.00014
2-[3-(3-fluorophenyl)-1H-indazol-1-yl]-1-(piperidin-1-yl)ethanone
Trypanosoma brucei
-
pH 8.0, 22C
0.1
3,3-dimethyl-1-(3-(1-methyl-1H-indol-4-yl)-1H-indazol-1-yl)butan-2-one
Trypanosoma brucei
-
above, pH and temperature not specified in the publication
0.1
3,3-dimethyl-1-(3-(naphthalen-2-yl)-1H-indazol-1-yl)butan-2-one
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0003
3,3-dimethyl-1-(3-(pyridin-4-yl)-1H-indazol-1-yl)butan-2-one
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0004
3,3-dimethyl-1-(3-(thiophen-3-yl)-1H-indazol-1-yl)butan-2-one
Trypanosoma brucei
-
above, pH and temperature not specified in the publication
0.0088
3,3-dimethyl-1-(5-phenyl-2H-tetrazol-2-yl)butan-2-one
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0035
3,3-dimethyl-1-[5-[3-(trifluoromethyl)phenyl]-2H-tetrazol-2-yl]butan-2-one
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.1
3,3-dimethyl-1-[5-[4-(trifluoromethyl)phenyl]-2H-tetrazol-2-yl]butan-2-one
Trypanosoma brucei
-
above, pH and temperature not specified in the publication
0.0058
3,5-dichloro-N-[(2-phenyl-1,3-thiazol-4-yl)methyl]benzenesulfonamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.1
3-((3-(3-fluorophenyl)-1H-indazol-1-yl)methyl)-5-methylisoxazole
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.1
3-((3-(3-fluorophenyl)-1H-indazol-1-yl)methyl)-5-phenylisoxazole
Trypanosoma brucei
-
above, pH and temperature not specified in the publication
0.0029
3-(3-fluorophenyl)-1-(pyridin-2-ylmethyl)-1H-indazole
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0045
3-(3-fluorophenyl)-1-(pyridin-3-ylmethyl)-1H-indazole
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.00008
3-(tert-butylsulfonylmethyl)-1-(3-fluorophenyl)-1H-indazole
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.00022
3-(tert-butylsulfonylmethyl)-1-phenyl-1H-indazole
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.028
4-((3-(3-fluorophenyl)-1H-indazol-1-yl)methyl)-2,5-dimethyloxazole
Trypanosoma brucei
-
above, pH and temperature not specified in the publication
0.0087
4-acetyl-N-[(2-phenyl-1,3-thiazol-4-yl)methyl]benzenesulfonamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.011
4-methoxy-N-[(3-phenyl-1,2,4-oxadiazol-5-yl)methyl]benzenesulfonamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.004
4-methoxy-N-[[3-(2-methylphenyl)-1,2,4-oxadiazol-5-yl]methyl]benzenesulfonamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.1
4-methyl-N-[(2-phenyl-1,3-thiazol-4-yl)methyl]benzenesulfonamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0117
betulin
Leishmania donovani
G5D5D5
pH 8.0, 25C, recombinant enzyme
0.0134
conessine
Leishmania donovani
G5D5D5
pH 8.0, 25C, recombinant enzyme
0.02
methyl (3-phenyl-1H-indazol-1-yl)acetate
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.1
N,N-dibenzyl-2-[3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl]acetamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0002
N,N-diethyl-2-(3-(3-fluorophenyl)-1H-indazol-1-yl)acetamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.015
N,N-diethyl-2-(3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl)acetamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0056
N-(1-phenylethyl)-2-[3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl]acetamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.000145
N-(2-(dimethylamino)ethyl)-2-(3-(3-fluorophenyl)-1H-indazol-1-yl)acetamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.03
N-(2-methoxyethyl)-2-[3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl]acetamide
Trypanosoma brucei
-
above, pH and temperature not specified in the publication
0.005
N-(2-phenylethyl)-2-[3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl]acetamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.00035
N-(3-(1H-imidazol-1-yl)propyl)-2-(3-(3-fluorophenyl)-1H-indazol-1-yl)acetamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.000045
N-(3-(dimethylamino)propyl)-2-(3-(3-fluorophenyl)-1H-indazol-1-yl)acetamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.1
N-(4-chlorobenzyl)-2-(3-(3-fluorophenyl)-1H-indazol-1-yl)acetamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.003
N-(4-chlorobenzyl)-2-(3-(trifluoromethyl)-4,5,6,7-tetrahydro-1Hindazol-1-yl)acetamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.05
N-(4-[methyl[(3-phenyl-1,2,4-oxadiazol-5-yl)methyl]sulfamoyl]phenyl)acetamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0059
N-(4-[[(3-phenyl-1,2,4-oxadiazol-5-yl)methyl]sulfamoyl]phenyl)acetamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.012
N-(butan-2-yl)-2-[3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl]acetamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0039
N-(thiophen-2-ylmethyl)-2-[3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl]acetamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.1
N-cyclohexyl-2-[3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl]acetamide
Trypanosoma brucei
-
above, pH and temperature not specified in the publication
0.00017
N-methyl-(3-(dimethylamino)propyl)-2-(3-(3-fluorophenyl)-1H-indazol-1-yl)acetamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.1
N-phenyl-2-[3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl]acetamide
Trypanosoma brucei
-
above, pH and temperature not specified in the publication
0.013
N-propyl-2-[3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl]acetamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.002 - 4
N-[(2-phenyl-1,3-thiazol-4-yl)methyl]naphthalene-2-sulfonamide
Trypanosoma brucei
-
above, pH and temperature not specified in the publication
0.05
N-[(3-phenyl-1,2,4-oxadiazol-5-yl)methyl]benzenesulfonamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.1
N-[(5-methyl-2-phenyl-2H-1,2,3-triazol-4-yl)methyl]naphthalene-1-sulfonamide
Trypanosoma brucei
-
above, pH and temperature not specified in the publication
0.0085
N-[(5-methyl-2-phenyl-2H-1,2,3-triazol-4-yl)methyl]thiophene-2-sulfonamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0037
N-[4-([[3-(2-chlorophenyl)-1,2,4-oxadiazol-5-yl]methyl]sulfamoyl)phenyl]acetamide
Trypanosoma brucei
-
above, pH and temperature not specified in the publication
0.0017
N-[4-([[3-(2-methylphenyl)-1,2,4-oxadiazol-5-yl]methyl]sulfamoyl)phenyl]acetamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.025
N-[[3-(2-chlorophenyl)-1,2,4-oxadiazol-5-yl]methyl]benzenesulfonamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.015
N-[[3-(2-methylphenyl)-1,2,4-oxadiazol-5-yl]methyl]benzenesulfonamide
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.0029
tert-butyl-4-(2-(2-(3-(3-fluorophenyl)-1H-indazol-1-yl)acetamido)ethyl)piperazine-1-carboxylate
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.018
tomatine
Leishmania donovani
G5D5D5
pH 8.0, 25C, recombinant enzyme
0.0112
uvaol
Leishmania donovani
G5D5D5
pH 8.0, 25C, recombinant enzyme
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2.1 - 2.2
purified recombinant trypanothione synthase performing the trypanothione synthase reaction, average of the activity of recombinant enzymes expressed from different constructs, overview
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 8
-
reaction step 1
7
-
reaction step 2, no activity at pH 8.0
7.2 - 7.5
assay at
7.5 - 7.8
-
synthesis of both mono-glutathionylspermidine and di-glutathionylspermidine conjugates
8.5
with substrates glutathionylspermidine and spermidine
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 11
inactive above or below
5.5 - 10
approx. 50% of maximal activity at pH 6.6 and pH 9.5 respectively
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 37
-
assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
10 - 60
residual activity above
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
64000
gel filtration
73560
deduced from nucleotide sequence
73740
MALDI mass spectrometry
73744
1 * 73744, MALDI mass spectrometry
81000
-
gel filtration
82000
-
x * 82000, SDS-PAGE
87000
-
1 * 87000, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
determination of three crystal forms, 18C, hanging drop vapor diffusion method, 0.001 ml of protein solution is mixed with 0.001 ml of reservoir solution, for crystal form I were optimized to a reservoir of 1.4 M (NH4)2SO4, 100 mM HEPES, pH 7.0, 200 mM NaBr, crystal form II to 1.6 M (NH4)2SO4, 100 mM HEPES, pH 7.0, and 200 mM NaBr, and crystal form III to 14% polyethylene glycol 8000, 15% glycerol and 100 mM KCl, within 2 days, X-ray diffraction structure determination and analysis at 2.3-3.6 A resolution, modelling; hanging gamma-drop vapor diffusion method, structure of Leishmania major trypanothione synthetase-amidase, determined in three crystal forms, reveals two catalytic domains
-
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
the C-terminal segment of the protein contributes to the amidase domain structure, and its loss likely destabilizes the fold
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate, DEAE-cellulose, partially purified
recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography, dialysis, and ion exchange chromatography
recombinant His-tagged enzyme from Escherichia coli strain BL21 star (DE3) by nickel affinity chromatography and tag cleavage through thrombin
recombinant His-tagged enzyme from Escherichia coli strain Tuner(DE3) by nickel affinity chromatography and tag cleavage by tobacco etch virus protease, followed by a second nickel affinity chromatography
-
recombinant His-tagged TcTryS, Ni2+-affinity, anion exchange
recombinant His-tagged wild-type enzyme and mutants from Escherichia coli by nickel affinity chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA sequence determination and analysis, expression of wild-type enzyme and mutants in Escherichia coli as His-tagged or Nus-fusion proteins
DNA sequence determination, cloning and RNAi transfection, expression in Escherichia coli
-
enzyme expression analysis, cloning and transfection of RNAi
-
expressed in Escherichia coli
-
expression in Escherichia coli
-
expression in Escherichia coli and Saccharomyces cerevisiae
expression of His-tagged enzyme in Escherichia coli strain BL21 star (DE3)
gene TRYS, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression of His-tagged enzyme in Escherichia coli
recombinant expression of His-tagged enzyme in Escherichia coli strain Tuner(DE3)
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C59A
-
mutant lacks the amidase activity, while the synthetase activity is similar to wild-type. Constructed for the production of trypanothione and trypanothione disulfide in >200 mg quantities. The protocol also allows the synthesis of related glutathione conjugates
R553E
site-directed mutagenesis
R553K
site-directed mutagenesis
R553L
site-directed mutagenesis
R553Q
site-directed mutagenesis
R613E
site-directed mutagenesis
R613K
site-directed mutagenesis
R613L
site-directed mutagenesis
R613Q
site-directed mutagenesis
DELTAN236
-
unable to produce trypanothione
DELTAN250
-
unable to produce trypanothione
C57A
-
functional synthetase devoid of amidase activity. In the presence of inducer, this line showed decreased growth in vitro and decreased virulence in vivo, indicating that the amidase function is not absolutely required for viability
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
drug development
pharmacology
-
the enzyme would serve as a potential target for antiparasitic chemotherapy
synthesis
-
production of trypanothione and trypanothione disulfide in >200 mg quantities by mutant C59A lacking the amidase activity. The protocol also allows the synthesis of related glutathione conjugates