Cloned (Comment) | Organism |
---|---|
recombinant expression of His-tagged enzyme in Escherichia coli strain Tuner(DE3) | Trypanosoma brucei |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
glutathione | substrate inhibition | Trypanosoma brucei | |
trypanothione | competitive feedback inhibition | Trypanosoma brucei |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0066 | - |
ATP | recombinant enzyme, pH 8.0, 25°C, HEPES buffer, with spermidine | Trypanosoma brucei | |
0.01 | - |
ATP | recombinant enzyme, pH 8.0, 37°C, HEPES buffer, with spermidine | Trypanosoma brucei | |
0.012 | - |
ATP | recombinant enzyme, pH 7.0, 37°C, phosphate buffer, with glutathionylspermidine | Trypanosoma brucei | |
0.012 | - |
N1-glutathionylspermidine | recombinant enzyme, pH 8.0, 25°C, HEPES buffer | Trypanosoma brucei | |
0.012 | - |
N8-glutathionylspermidine | recombinant enzyme, pH 8.0, 25°C, HEPES buffer | Trypanosoma brucei | |
0.016 | - |
N1-glutathionylspermidine | recombinant enzyme, pH 8.0, 37°C, HEPES buffer | Trypanosoma brucei | |
0.016 | - |
N8-glutathionylspermidine | recombinant enzyme, pH 8.0, 37°C, HEPES buffer | Trypanosoma brucei | |
0.018 | - |
ATP | recombinant enzyme, pH 7.0, 37°C, phosphate buffer, with spermidine | Trypanosoma brucei | |
0.032 | - |
N1-glutathionylspermidine | recombinant enzyme, pH 7.0, 37°C, phosphate buffer | Trypanosoma brucei | |
0.032 | - |
N8-glutathionylspermidine | recombinant enzyme, pH 7.0, 37°C, phosphate buffer | Trypanosoma brucei | |
0.032 | - |
glutathione | recombinant enzyme, pH 8.0, 25°C, HEPES buffer, with spermidine | Trypanosoma brucei | |
0.034 | - |
glutathione | recombinant enzyme, pH 7.0, 37°C, phosphate buffer, with glutathionylspermidine | Trypanosoma brucei | |
0.045 | - |
glutathione | recombinant enzyme, pH 8.0, 37°C, HEPES buffer, with spermidine | Trypanosoma brucei | |
0.069 | - |
glutathione | recombinant enzyme, pH 7.0, 37°C, phosphate buffer, with spermidine | Trypanosoma brucei | |
0.092 | - |
spermidine | recombinant enzyme, pH 8.0, 25°C, HEPES buffer | Trypanosoma brucei | |
0.139 | - |
spermidine | recombinant enzyme, pH 8.0, 37°C, HEPES buffer | Trypanosoma brucei | |
0.687 | - |
spermidine | recombinant enzyme, pH 7.0, 37°C, phosphate buffer | Trypanosoma brucei |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Trypanosoma brucei |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 glutathione + spermidine + 2 ATP | Trypanosoma brucei | 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 | N1,N8-bis(glutathionyl)spermidine + 2 ADP + 2 phosphate | - |
? | |
2 glutathione + spermidine + 2 ATP | Trypanosoma brucei 449 | 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 | N1,N8-bis(glutathionyl)spermidine + 2 ADP + 2 phosphate | - |
? | |
glutathione + spermidine + ATP | Trypanosoma brucei | reaction of EC 6.3.1.8, step one | N1-glutathionylspermidine + N8-glutathionylspermidine + ADP + phosphate | - |
? | |
glutathione + spermidine + ATP | Trypanosoma brucei 449 | reaction of EC 6.3.1.8, step one | N1-glutathionylspermidine + N8-glutathionylspermidine + ADP + phosphate | - |
? | |
additional information | Trypanosoma brucei | in pathogenic trypanosomes, trypanothione synthetase catalyzes the synthesis of both glutathionylspermidine and trypanothione (bis(glutathionyl)spermidine). Trypanothione is synthesized from two molecules of GSH that are covalently bridged by a spermidine in two ATP-dependent reactions. The enzyme also shows trypanothione amidase activity catalyzed at a second distinct active site | ? | - |
? | |
additional information | Trypanosoma brucei 449 | in pathogenic trypanosomes, trypanothione synthetase catalyzes the synthesis of both glutathionylspermidine and trypanothione (bis(glutathionyl)spermidine). Trypanothione is synthesized from two molecules of GSH that are covalently bridged by a spermidine in two ATP-dependent reactions. The enzyme also shows trypanothione amidase activity catalyzed at a second distinct active site | ? | - |
? | |
N1-glutathionylspermidine + glutathione + ATP | Trypanosoma brucei | step two | N1,N8-bis(glutathionyl)spermidine + ADP + phosphate | - |
? | |
N1-glutathionylspermidine + glutathione + ATP | Trypanosoma brucei 449 | step two | N1,N8-bis(glutathionyl)spermidine + ADP + phosphate | - |
? | |
N8-glutathionylspermidine + glutathione + ATP | Trypanosoma brucei | step two | N1,N8-bis(glutathionyl)spermidine + ADP + phosphate | - |
? | |
N8-glutathionylspermidine + glutathione + ATP | Trypanosoma brucei 449 | step two | N1,N8-bis(glutathionyl)spermidine + ADP + phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Trypanosoma brucei | - |
- |
- |
Trypanosoma brucei 449 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
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 | Trypanosoma brucei |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
glutathione + spermidine + ATP = glutathionylspermidine + ADP + phosphate | ter-reactant mechanism of TryS catalysis, modeling, overview | Trypanosoma brucei |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 glutathione + spermidine + 2 ATP | 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 | Trypanosoma brucei | N1,N8-bis(glutathionyl)spermidine + 2 ADP + 2 phosphate | - |
? | |
2 glutathione + spermidine + 2 ATP | 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 | Trypanosoma brucei 449 | N1,N8-bis(glutathionyl)spermidine + 2 ADP + 2 phosphate | - |
? | |
glutathione + spermidine + ATP | reaction of EC 6.3.1.8, step one | Trypanosoma brucei | N1-glutathionylspermidine + N8-glutathionylspermidine + ADP + phosphate | - |
? | |
glutathione + spermidine + ATP | reaction of EC 6.3.1.8, step one | Trypanosoma brucei 449 | N1-glutathionylspermidine + N8-glutathionylspermidine + ADP + phosphate | - |
? | |
additional information | in pathogenic trypanosomes, trypanothione synthetase catalyzes the synthesis of both glutathionylspermidine and trypanothione (bis(glutathionyl)spermidine). Trypanothione is synthesized from two molecules of GSH that are covalently bridged by a spermidine in two ATP-dependent reactions. The enzyme also shows trypanothione amidase activity catalyzed at a second distinct active site | Trypanosoma brucei | ? | - |
? | |
additional information | in pathogenic trypanosomes, trypanothione synthetase catalyzes the synthesis of both glutathionylspermidine and trypanothione (bis(glutathionyl)spermidine). Trypanothione is synthesized from two molecules of GSH that are covalently bridged by a spermidine in two ATP-dependent reactions. The enzyme also shows trypanothione amidase activity catalyzed at a second distinct active site | Trypanosoma brucei 449 | ? | - |
? | |
N1-glutathionylspermidine + glutathione + ATP | step two | Trypanosoma brucei | N1,N8-bis(glutathionyl)spermidine + ADP + phosphate | - |
? | |
N1-glutathionylspermidine + glutathione + ATP | step two | Trypanosoma brucei 449 | N1,N8-bis(glutathionyl)spermidine + ADP + phosphate | - |
? | |
N8-glutathionylspermidine + glutathione + ATP | step two | Trypanosoma brucei | N1,N8-bis(glutathionyl)spermidine + ADP + phosphate | - |
? | |
N8-glutathionylspermidine + glutathione + ATP | step two | Trypanosoma brucei 449 | N1,N8-bis(glutathionyl)spermidine + ADP + phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Trypanothione synthetase | - |
Trypanosoma brucei |
TryS | - |
Trypanosoma brucei |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | 37 | assay at | Trypanosoma brucei |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.8 | - |
glutathione | recombinant enzyme, pH 8.0, 25°C, HEPES buffer, with spermidine | Trypanosoma brucei | |
2.1 | - |
glutathione | recombinant enzyme, pH 7.0, 37°C, phosphate buffer, with glutathionylspermidine | Trypanosoma brucei | |
2.8 | - |
glutathione | recombinant enzyme, pH 7.0, 37°C, phosphate buffer, with spermidine | Trypanosoma brucei | |
5.2 | - |
glutathione | recombinant enzyme, pH 8.0, 37°C, HEPES buffer, with spermidine | Trypanosoma brucei |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | 8 | reaction step 1 | Trypanosoma brucei |
7 | - |
reaction step 2, no activity at pH 8.0 | Trypanosoma brucei |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Trypanosoma brucei |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.132 | - |
glutathione | recombinant enzyme, pH 8.0, 25°C, HEPES buffer, with spermidine | Trypanosoma brucei | |
0.223 | - |
trypanothione | recombinant enzyme, pH 7.0, 37°C, phosphate buffer, with glutathione as substrate, versus spermidine | Trypanosoma brucei | |
0.228 | - |
trypanothione | recombinant enzyme, pH 7.0, 37°C, phosphate buffer, with spermidine as substrate, versus spermidine | Trypanosoma brucei | |
0.332 | - |
glutathione | recombinant enzyme, pH 8.0, 37°C, HEPES buffer, with spermidine | Trypanosoma brucei | |
0.346 | - |
trypanothione | recombinant enzyme, pH 7.0, 37°C, phosphate buffer, with glutathione as substrate, versus glutathione | Trypanosoma brucei | |
0.377 | - |
trypanothione | recombinant enzyme, pH 7.0, 37°C, phosphate buffer, with spermidine as substrate, versus glutathione | Trypanosoma brucei | |
0.849 | - |
glutathione | recombinant enzyme, pH 7.0, 37°C, phosphate buffer, with spermidine | Trypanosoma brucei | |
1.085 | - |
glutathione | recombinant enzyme, pH 7.0, 37°C, phosphate buffer, versus glutathionylspermidine | Trypanosoma brucei |
General Information | Comment | Organism |
---|---|---|
additional information | the steady-state kinetic analysis discard a ping-pong mechanism | Trypanosoma brucei |