Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli WK6 cells | Trypanosoma vivax |
Crystallization (Comment) | Organism |
---|---|
in complex with immucillin H, hanging drop vapour diffusion method, with 20% polyethylene glycol 4000 and 0.25 M ammonium sulfate in 10 mM potassium acetate, pH 5.0 | Trypanosoma vivax |
Protein Variants | Comment | Organism |
---|---|---|
C248P | mutant shows decreased kcat value compared to the wild type enzyme using guanosine as substrate | Trypanosoma vivax |
D253P | mutant shows increased kcat value compared to the wild type enzyme using guanosine as substrate | Trypanosoma vivax |
E249P | mutant shows decreased kcat value compared to the wild type enzyme using guanosine as substrate | Trypanosoma vivax |
H247P | mutant shows increased kcat value compared to the wild type enzyme using guanosine as substrate | Trypanosoma vivax |
L250P | mutant shows decreased kcat value compared to the wild type enzyme using guanosine as substrate | Trypanosoma vivax |
L251P | mutant shows increased kcat value compared to the wild type enzyme using guanosine as substrate | Trypanosoma vivax |
additional information | the loop deletion mutant 3G hydrolyzes guanosine and 2-aminopurine riboside with a turnover comparable to that of the wild type, but an increase in Km is observed for both substrates, 7-methyl guanosine is hydrolyzed even faster by 3G compared with the wild type and hardly any difference is observed in the Km | Trypanosoma vivax |
R252P | mutant shows increased kcat value compared to the wild type enzyme using guanosine as substrate | Trypanosoma vivax |
General Stability | Organism |
---|---|
once thawed, enzyme stocks are kept at 4°C and used over a period of several days but never refrozen to avoid activity loss | Trypanosoma vivax |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
immucillin H | immucillin H is (1S)-1-(9-deazaadenin-9-yl)-1,4-dideoxy-1,4-imino-D-ribitol | Trypanosoma vivax |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0005 | - |
7-methyl guanosine | wild type enzyme, in 50 mM potassium phosphate buffer, at 25°C and pH 7.0 | Trypanosoma vivax | |
0.0018 | - |
guanosine | mutant enzyme D253P, in 50 mM potassium phosphate buffer, at 25°C and pH 7.0 | Trypanosoma vivax | |
0.0023 | - |
guanosine | wild type enzyme, in 50 mM potassium phosphate buffer, at 25°C and pH 7.0 | Trypanosoma vivax | |
0.003 | - |
guanosine | mutant enzyme E249P, in 50 mM potassium phosphate buffer, at 25°C and pH 7.0 | Trypanosoma vivax | |
0.004 | - |
guanosine | mutant enzyme C248P, in 50 mM potassium phosphate buffer, at 25°C and pH 7.0 | Trypanosoma vivax | |
0.008 | - |
guanosine | mutant enzyme H247P, in 50 mM potassium phosphate buffer, at 25°C and pH 7.0 | Trypanosoma vivax | |
0.009 | - |
2-aminopurine riboside | wild type enzyme, in 50 mM potassium phosphate buffer, at 25°C and pH 7.0 | Trypanosoma vivax | |
0.016 | - |
guanosine | mutant enzyme L250P, in 50 mM potassium phosphate buffer, at 25°C and pH 7.0 | Trypanosoma vivax | |
0.021 | - |
guanosine | mutant enzyme L251P, in 50 mM potassium phosphate buffer, at 25°C and pH 7.0 | Trypanosoma vivax | |
0.024 | - |
guanosine | mutant enzyme R252P, in 50 mM potassium phosphate buffer, at 25°C and pH 7.0 | Trypanosoma vivax |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Trypanosoma vivax | Q9GPQ4 | - |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA column chromatography and Superdex-75 gel filtration | Trypanosoma vivax |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-aminopurine riboside + H2O | - |
Trypanosoma vivax | 2-aminopurine + D-ribose | - |
? | |
7-methyl guanosine + H2O | - |
Trypanosoma vivax | D-ribose + 7-methyl guanine | - |
? | |
adenosine + H2O | - |
Trypanosoma vivax | D-ribose + adenine | - |
? | |
guanosine + H2O | - |
Trypanosoma vivax | D-ribose + guanine | - |
? | |
inosine + H2O | - |
Trypanosoma vivax | D-ribose + hypoxanthine | - |
? |
Synonyms | Comment | Organism |
---|---|---|
purine-specific nucleoside hydrolase | - |
Trypanosoma vivax |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.57 | - |
2-aminopurine riboside | wild type enzyme, in 50 mM potassium phosphate buffer, at 25°C and pH 7.0 | Trypanosoma vivax | |
1.07 | - |
7-methyl guanosine | wild type enzyme, in 50 mM potassium phosphate buffer, at 25°C and pH 7.0 | Trypanosoma vivax | |
1.09 | - |
guanosine | mutant enzyme C248P, in 50 mM potassium phosphate buffer, at 25°C and pH 7.0 | Trypanosoma vivax | |
1.4 | - |
guanosine | mutant enzyme L250P, in 50 mM potassium phosphate buffer, at 25°C and pH 7.0 | Trypanosoma vivax | |
1.7 | - |
guanosine | mutant enzyme E249P, in 50 mM potassium phosphate buffer, at 25°C and pH 7.0 | Trypanosoma vivax | |
2.3 | - |
guanosine | wild type enzyme, in 50 mM potassium phosphate buffer, at 25°C and pH 7.0 | Trypanosoma vivax | |
2.5 | - |
guanosine | mutant enzyme L251P, in 50 mM potassium phosphate buffer, at 25°C and pH 7.0 | Trypanosoma vivax | |
3.9 | - |
guanosine | mutant enzyme D253P, in 50 mM potassium phosphate buffer, at 25°C and pH 7.0 | Trypanosoma vivax | |
4.1 | - |
guanosine | mutant enzyme H247P, in 50 mM potassium phosphate buffer, at 25°C and pH 7.0 | Trypanosoma vivax | |
13 | - |
guanosine | mutant enzyme R252P, in 50 mM potassium phosphate buffer, at 25°C and pH 7.0 | Trypanosoma vivax |