BRENDA - Enzyme Database

Site-directed mutagenesis of the redox-active cysteines of Trypanosoma cruzi trypanothione reductase

Borges, A.; Cunningham, M.L.; Tovar, J.; Fairlamb, A.H.; Eur. J. Biochem. 228, 745-752 (1995) View publication on PubMed

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
1.8.1.12
additional information
activity is dependent on a redox-active cysteine at the active centre
Trypanosoma cruzi
Cloned(Commentary)
EC Number
Cloned (Commentary)
Organism
1.8.1.12
overexpression of wild-type and mutants C53S, C53A, C58S in Escherichia coli
Trypanosoma cruzi
Engineering
EC Number
Protein Variants
Commentary
Organism
1.8.1.12
C53A
site-directed mutagenesis, without catalytic active cysteine residue in the active center, no activity with trypanothione disulfide and NADPH, but showing transhydrogenase activity between NADPH and thio-NADP+
Trypanosoma cruzi
1.8.1.12
C53S
site-directed mutagenesis, without catalytic active cysteine residue in the active center, no activity with trypanothione disulfide and NADPH, but showing transhydrogenase activity between NADPH and thio-NADP+
Trypanosoma cruzi
1.8.1.12
C58S
site-directed mutagenesis, without catalytic active cysteine residue in the active center, no activity with trypanothione disulfide and NADPH, but showing transhydrogenase activity between NADPH and thio-NADP+
Trypanosoma cruzi
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.8.1.12
0.004
0.015
thio-NADP+
recombinant mutants C53S, C53A, C58S
Trypanosoma cruzi
1.8.1.12
0.007
0.014
NADPH
recombinant mutants C53S, C53A, C58S
Trypanosoma cruzi
1.8.1.12
0.017
-
NADPH
recombinant wild-type
Trypanosoma cruzi
1.8.1.12
0.024
-
trypanothione disulfide
recombinant wild-type
Trypanosoma cruzi
1.8.1.12
0.046
-
glutathionylspermidine disulfide
recombinant wild-type
Trypanosoma cruzi
Organism
EC Number
Organism
UniProt
Commentary
Textmining
1.8.1.12
Trypanosoma cruzi
-
Silvio strain
-
1.8.1.12
Trypanosoma cruzi Silvio
-
Silvio strain
-
Purification (Commentary)
EC Number
Purification (Commentary)
Organism
1.8.1.12
recombinant wild-type and mutants C53S, C53A, C58S from Escherichia coli
Trypanosoma cruzi
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.8.1.12
0.18
-
recombinant mutant C58S: transhydrogenase activity between NADPH and thio-NADP+
Trypanosoma cruzi
1.8.1.12
0.21
-
recombinant mutant C53A: transhydrogenase activity between NADPH and thio-NADP+
Trypanosoma cruzi
1.8.1.12
0.44
-
recombinant mutant C53S: transhydrogenase activity between NADPH and thio-NADP+
Trypanosoma cruzi
1.8.1.12
6.49
-
recombinant wild-type: transhydrogenase activity between NADPH and thio-NADP+
Trypanosoma cruzi
1.8.1.12
143.2
-
purified recombinant enzyme
Trypanosoma cruzi
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
1.8.1.12
glutathionylspermidine disulfide + NADPH
-
394800
Trypanosoma cruzi
2-glutathionylspermidine + NADP+
-
-
-
?
1.8.1.12
glutathionylspermidine disulfide + NADPH
-
394800
Trypanosoma cruzi Silvio
2-glutathionylspermidine + NADP+
-
-
-
?
1.8.1.12
additional information
wild-type enzyme shows transhydrogenase activity between NADPH and thio-NADP+
394800
Trypanosoma cruzi
?
-
-
-
?
1.8.1.12
additional information
recombinant mutants C53S, C53A, C58S: no activity with trypanothione disulfide and NADPH, but showing transhydrogenase activity between NADPH and thio-NADP+
394800
Trypanosoma cruzi
?
-
-
-
?
1.8.1.12
additional information
wild-type enzyme shows transhydrogenase activity between NADPH and thio-NADP+
394800
Trypanosoma cruzi Silvio
?
-
-
-
?
1.8.1.12
additional information
recombinant mutants C53S, C53A, C58S: no activity with trypanothione disulfide and NADPH, but showing transhydrogenase activity between NADPH and thio-NADP+
394800
Trypanosoma cruzi Silvio
?
-
-
-
?
1.8.1.12
NADPH + H+ + thio-NADP+
-
394800
Trypanosoma cruzi
NADP+ + thio-NADPH + H+
-
-
-
r
1.8.1.12
NADPH + H+ + thio-NADP+
-
394800
Trypanosoma cruzi Silvio
NADP+ + thio-NADPH + H+
-
-
-
r
1.8.1.12
trypanothione disulfide + NADPH
-
394800
Trypanosoma cruzi
trypanothione + NADP+
-
394800
Trypanosoma cruzi
?
1.8.1.12
trypanothione disulfide + NADPH
-
394800
Trypanosoma cruzi Silvio
trypanothione + NADP+
-
394800
Trypanosoma cruzi Silvio
?
1.8.1.12
trypanothione disulfide + thio-NADPH + H+
-
394800
Trypanosoma cruzi
trypanothione + thio-NADP+
-
-
-
r
Synonyms
EC Number
Synonyms
Commentary
Organism
1.8.1.12
TR
-
Trypanosoma cruzi
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.8.1.12
25
-
assay at
Trypanosoma cruzi
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.8.1.12
0.19
0.47
thio-NADPH
recombinant mutants C53S, C53A, C58S
Trypanosoma cruzi
1.8.1.12
0.21
0.51
NADPH
recombinant mutants C53S, C53A, C58S
Trypanosoma cruzi
1.8.1.12
6.33
-
NADPH
recombinant wild-type
Trypanosoma cruzi
1.8.1.12
7.02
-
thio-NADPH
recombinant wild-type
Trypanosoma cruzi
1.8.1.12
85.4
-
glutathionylspermidine disulfide
recombinant wild-type
Trypanosoma cruzi
1.8.1.12
108
-
trypanothione disulfide
recombinant wild-type
Trypanosoma cruzi
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.8.1.12
beta-thio-NADPH
-
Trypanosoma cruzi
1.8.1.12
FAD
-
Trypanosoma cruzi
1.8.1.12
NADPH
-
Trypanosoma cruzi
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
1.8.1.12
additional information
activity is dependent on a redox-active cysteine at the active centre
Trypanosoma cruzi
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.8.1.12
overexpression of wild-type and mutants C53S, C53A, C58S in Escherichia coli
Trypanosoma cruzi
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.8.1.12
beta-thio-NADPH
-
Trypanosoma cruzi
1.8.1.12
FAD
-
Trypanosoma cruzi
1.8.1.12
NADPH
-
Trypanosoma cruzi
Engineering (protein specific)
EC Number
Protein Variants
Commentary
Organism
1.8.1.12
C53A
site-directed mutagenesis, without catalytic active cysteine residue in the active center, no activity with trypanothione disulfide and NADPH, but showing transhydrogenase activity between NADPH and thio-NADP+
Trypanosoma cruzi
1.8.1.12
C53S
site-directed mutagenesis, without catalytic active cysteine residue in the active center, no activity with trypanothione disulfide and NADPH, but showing transhydrogenase activity between NADPH and thio-NADP+
Trypanosoma cruzi
1.8.1.12
C58S
site-directed mutagenesis, without catalytic active cysteine residue in the active center, no activity with trypanothione disulfide and NADPH, but showing transhydrogenase activity between NADPH and thio-NADP+
Trypanosoma cruzi
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.8.1.12
0.004
0.015
thio-NADP+
recombinant mutants C53S, C53A, C58S
Trypanosoma cruzi
1.8.1.12
0.007
0.014
NADPH
recombinant mutants C53S, C53A, C58S
Trypanosoma cruzi
1.8.1.12
0.017
-
NADPH
recombinant wild-type
Trypanosoma cruzi
1.8.1.12
0.024
-
trypanothione disulfide
recombinant wild-type
Trypanosoma cruzi
1.8.1.12
0.046
-
glutathionylspermidine disulfide
recombinant wild-type
Trypanosoma cruzi
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.8.1.12
recombinant wild-type and mutants C53S, C53A, C58S from Escherichia coli
Trypanosoma cruzi
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.8.1.12
0.18
-
recombinant mutant C58S: transhydrogenase activity between NADPH and thio-NADP+
Trypanosoma cruzi
1.8.1.12
0.21
-
recombinant mutant C53A: transhydrogenase activity between NADPH and thio-NADP+
Trypanosoma cruzi
1.8.1.12
0.44
-
recombinant mutant C53S: transhydrogenase activity between NADPH and thio-NADP+
Trypanosoma cruzi
1.8.1.12
6.49
-
recombinant wild-type: transhydrogenase activity between NADPH and thio-NADP+
Trypanosoma cruzi
1.8.1.12
143.2
-
purified recombinant enzyme
Trypanosoma cruzi
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
1.8.1.12
glutathionylspermidine disulfide + NADPH
-
394800
Trypanosoma cruzi
2-glutathionylspermidine + NADP+
-
-
-
?
1.8.1.12
glutathionylspermidine disulfide + NADPH
-
394800
Trypanosoma cruzi Silvio
2-glutathionylspermidine + NADP+
-
-
-
?
1.8.1.12
additional information
wild-type enzyme shows transhydrogenase activity between NADPH and thio-NADP+
394800
Trypanosoma cruzi
?
-
-
-
?
1.8.1.12
additional information
recombinant mutants C53S, C53A, C58S: no activity with trypanothione disulfide and NADPH, but showing transhydrogenase activity between NADPH and thio-NADP+
394800
Trypanosoma cruzi
?
-
-
-
?
1.8.1.12
additional information
wild-type enzyme shows transhydrogenase activity between NADPH and thio-NADP+
394800
Trypanosoma cruzi Silvio
?
-
-
-
?
1.8.1.12
additional information
recombinant mutants C53S, C53A, C58S: no activity with trypanothione disulfide and NADPH, but showing transhydrogenase activity between NADPH and thio-NADP+
394800
Trypanosoma cruzi Silvio
?
-
-
-
?
1.8.1.12
NADPH + H+ + thio-NADP+
-
394800
Trypanosoma cruzi
NADP+ + thio-NADPH + H+
-
-
-
r
1.8.1.12
NADPH + H+ + thio-NADP+
-
394800
Trypanosoma cruzi Silvio
NADP+ + thio-NADPH + H+
-
-
-
r
1.8.1.12
trypanothione disulfide + NADPH
-
394800
Trypanosoma cruzi
trypanothione + NADP+
-
394800
Trypanosoma cruzi
?
1.8.1.12
trypanothione disulfide + NADPH
-
394800
Trypanosoma cruzi Silvio
trypanothione + NADP+
-
394800
Trypanosoma cruzi Silvio
?
1.8.1.12
trypanothione disulfide + thio-NADPH + H+
-
394800
Trypanosoma cruzi
trypanothione + thio-NADP+
-
-
-
r
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.8.1.12
25
-
assay at
Trypanosoma cruzi
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.8.1.12
0.19
0.47
thio-NADPH
recombinant mutants C53S, C53A, C58S
Trypanosoma cruzi
1.8.1.12
0.21
0.51
NADPH
recombinant mutants C53S, C53A, C58S
Trypanosoma cruzi
1.8.1.12
6.33
-
NADPH
recombinant wild-type
Trypanosoma cruzi
1.8.1.12
7.02
-
thio-NADPH
recombinant wild-type
Trypanosoma cruzi
1.8.1.12
85.4
-
glutathionylspermidine disulfide
recombinant wild-type
Trypanosoma cruzi
1.8.1.12
108
-
trypanothione disulfide
recombinant wild-type
Trypanosoma cruzi