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Information on EC 6.3.1.9 - trypanothione synthase and Organism(s) Leishmania major and UniProt Accession Q711P7
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6.3.1.9 trypanothione synthaseIUBMB Comments
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).
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Word Map
- 6.3.1.9
- trypanosoma
- leishmania
-
trypanosomatids
- brucei
- cruzi
- fasciculata
- crithidia
-
n1,n8-bisglutathionylspermidine
-
chagas
- infantum
-
tryparedoxine
-
antileishmanial
-
parasite-specific
- kinetoplastida
-
antitrypanosomal
- drug development
- synthesis
- pharmacology
The taxonomic range for the selected organisms is: Leishmania major
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
Reaction Schemes
Synonyms
trypanothione synthetase, tctrys, ldtrys, tsh synthetase, litrys, tbtrys, trypanothione synthase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
trypanothione synthetase-amidase
TryS
Cf-TS
-
-
-
-
GenBank AF006615-derived protein GI 3004644
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-
-
-
Synthetase, trypanothione
-
-
-
-
Synthetase, trypanothione (Crithidia fasciculata strain HS6 gene Cf-TS)
-
-
-
-
Trypanothione synthetase
Trypanothione synthetase (Crithidia fasciculata strain HS6 gene Cf-TS)
-
-
-
-
TSH synthetase
-
-
-
-
TryS
the enzyme has two domains: an ATP-dependent synthetase domain that generates the intermediate glutathionylspermidine and the final product trypanothione from glutathione and spermidine, and an amidase domain which can hydrolyse glutathionylspermidine and trypanothione to the original substrates
Trypanothione synthetase
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Acid amide formation
-
-
-
-
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
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)
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
2 glutathione + spermidine + 2 ATP
N1,N8-bis(glutathionyl)spermidine + 2 ADP + 2 phosphate
trypanothione synthase overall reaction
-
-
?
glutathione + glutathionylspermidine + ATP
N1,N8-bis(glutathionyl)spermidine + ADP + phosphate
2 glutathione + spermidine + 2 ATP
N1,N8-bis(glutathionyl)spermidine + 2 ADP + 2 phosphate
-
overall reaction, the enzyme catalyzes the formation of N1,N8-bis(glutathionyl)spermidine in two reaction steps, exhibiting activity of EC 6.3.1.8, glutathionylspermidine synthase, and 6.3.1.9, trypanothione synthase, overview
-
-
?
glutathione + spermidine + ATP
N1-glutathionylspermidine + N8-glutathionylspermidine + ADP + phosphate
-
reaction of EC 6.3.1.8, step one
-
-
?
N1-glutathionylspermidine + glutathione + ATP
N1,N8-bis(glutathionyl)spermidine + ADP + phosphate
-
step two
-
-
?
N8-glutathionylspermidine + glutathione + ATP
N1,N8-bis(glutathionyl)spermidine + ADP + phosphate
-
step two, N8-glutathionylspermidine is the favoured substrate
-
-
?
glutathione + glutathionylspermidine + ATP
N1,N8-bis(glutathionyl)spermidine + ADP + phosphate
trypanothione synthase catalyzes both the reaction of glutathionyspermidine synthase, EC 6.3.1.8, and of trypanothione synthase, EC 6.3.1.9
-
-
?
glutathione + glutathionylspermidine + ATP
N1,N8-bis(glutathionyl)spermidine + ADP + phosphate
trypanothione synthase catalyzes both the reaction of glutathionylspermidine synthase, EC 6.3.1.8, and of trypanothione synthase, EC 6.3.1.9
-
-
?
glutathione + spermidine + ATP
glutathionylspermidine + ADP + phosphate
-
-
-
?
glutathione + spermidine + ATP
glutathionylspermidine + ADP + phosphate
trypanothione synthase catalyzes both the reaction of glutathionyspermidine synthase, EC 6.3.1.8, and of trypanothione synthase, EC 6.3.1.9
-
-
?
glutathione + spermidine + ATP
glutathionylspermidine + ADP + phosphate
trypanothione synthase catalyzes both the reaction of glutathionylspermidine synthase, EC 6.3.1.8, and of trypanothione synthase, EC 6.3.1.9
-
-
?
glutathione + spermidine + ATP
glutathionylspermidine + ADP + phosphate
ATP in form of MgATP2-
-
-
?
additional information
?
-
phylogenetic analysis implies that TryS will replace the glutathionylspermidine synthase/trypanothione synthase complex in Leishmia major loosing the redundant GspS pseudogene from its genome
-
-
?
additional information
?
-
-
phylogenetic analysis implies that TryS will replace the glutathionylspermidine synthase/trypanothione synthase complex in Leishmia major loosing the redundant GspS pseudogene from its genome
-
-
?
additional information
?
-
the bifunctional trypanothione synthetase-amidase catalyzes biosynthesis and hydrolysis of the glutathione-spermidine adduct trypanothione, the principal intracellular thiolredox metabolite in parasitic trypanosomatids
-
-
?
additional information
?
-
structure modelling of substrate binding to the active site, overview
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
2 glutathione + spermidine + 2 ATP
N1,N8-bis(glutathionyl)spermidine + 2 ADP + 2 phosphate
trypanothione synthase overall reaction
-
-
?
glutathione + glutathionylspermidine + ATP
N1,N8-bis(glutathionyl)spermidine + ADP + phosphate
trypanothione synthase catalyzes both the reaction of glutathionyspermidine synthase, EC 6.3.1.8, and of trypanothione synthase, EC 6.3.1.9
-
-
?
glutathione + spermidine + ATP
glutathionylspermidine + ADP + phosphate
trypanothione synthase catalyzes both the reaction of glutathionyspermidine synthase, EC 6.3.1.8, and of trypanothione synthase, EC 6.3.1.9
-
-
?
2 glutathione + spermidine + 2 ATP
N1,N8-bis(glutathionyl)spermidine + 2 ADP + 2 phosphate
-
overall reaction, the enzyme catalyzes the formation of N1,N8-bis(glutathionyl)spermidine in two reaction steps, exhibiting activity of EC 6.3.1.8, glutathionylspermidine synthase, and 6.3.1.9, trypanothione synthase, overview
-
-
?
glutathione + spermidine + ATP
N1-glutathionylspermidine + N8-glutathionylspermidine + ADP + phosphate
-
reaction of EC 6.3.1.8, step one
-
-
?
N1-glutathionylspermidine + glutathione + ATP
N1,N8-bis(glutathionyl)spermidine + ADP + phosphate
-
step two
-
-
?
N8-glutathionylspermidine + glutathione + ATP
N1,N8-bis(glutathionyl)spermidine + ADP + phosphate
-
step two, N8-glutathionylspermidine is the favoured substrate
-
-
?
additional information
?
-
phylogenetic analysis implies that TryS will replace the glutathionylspermidine synthase/trypanothione synthase complex in Leishmia major loosing the redundant GspS pseudogene from its genome
-
-
?
additional information
?
-
-
phylogenetic analysis implies that TryS will replace the glutathionylspermidine synthase/trypanothione synthase complex in Leishmia major loosing the redundant GspS pseudogene from its genome
-
-
?
additional information
?
-
the bifunctional trypanothione synthetase-amidase catalyzes biosynthesis and hydrolysis of the glutathione-spermidine adduct trypanothione, the principal intracellular thiolredox metabolite in parasitic trypanosomatids
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
ATP
as MgATP2-, the enzyme is specific for ATP and shows only negligible activity with other NTPs
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
-
binding sites for N8-glutathionylspermidine , two Mg2+ ions, structure modeling, overview. For the second step of trypanothione synthesis glutathionylspermidine is bound in a way that preferentially allows N1-glutathionylation of N8-glutathionylspermidine, classifying N8-glutathionylspermidine as the favoured substrate
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.04
glutathionylspermidine
pH 7.7, 25°C, recombinant trypanothione synthase
additional information
additional information
hyperbolic kinetics, steady-state kinetics
-
additional information
additional information
-
hyperbolic kinetics, steady-state kinetics
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
metabolism
-
trypanothione synthetase catalyses the two-step biosynthesis of trypanothione from spermidine and glutathione
additional information
-
binding sites for glutathione, ATP and two Mg2+ ions, molecular docking and molecular dynamics simulations, strructure modeling, detailed overview
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). Q711P7_LEIMA
652
0
74434
TrEMBL
other Location (Reliability: 5)
PDB
SCOP
CATH
UNIPROT
ORGANISM
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
secondary, tertiary, and domain structures, determined from three crystal forms, revealing two catalytic domains. The N-terminal domain, a cysteine, histidine-dependent amidohydrolase/peptidase amidase, is a papain-like cysteine protease, and the C-terminal synthetase domain displays an ATP-grasp family fold common to C:N ligases
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
determination of three crystal forms, 18°C, 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
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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
recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography, dialysis, and ion exchange chromatography
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene TRYS, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression of His-tagged enzyme in Escherichia coli
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Oza, S.L.; Shaw, M.P.; Wyllie, S.; Fairlamb, A.H.
Trypanothione biosynthesis in Leishmania major
Mol. Biochem. Parasitol.
139
107-116
2005
Leishmania major (Q711P7), Leishmania major
Oza, S.L.; Wyllie, S.; Fairlamb, A.H.
Mapping the functional synthetase domain of trypanothione synthetase from Leishmania major
Mol. Biochem. Parasitol.
149
117-120
2006
Leishmania major (Q711P7), Leishmania major
Fyfe, P.K.; Oza, S.L.; Fairlamb, A.H.; Hunter, W.N.
Leishmania trypanothione synthetase-amidase structure reveals a basis for regulation of conflicting synthetic and hydrolytic activities
J. Biol. Chem.
283
17672-17680
2008
Leishmania major (Q711P7)
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