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Literature summary extracted from
- Structural basis for substrate specificity in group I nucleoside hydrolases (2008), Biochemistry, 47, 4418-4426.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 3.2.2.8 | expressed in Escherichia coli BL21(DE3) cells | Escherichia coli |
Crystallization (Commentary)
| EC Number | Crystallization (Comment) | Organism |
|---|---|---|
| 3.2.2.8 | hanging drop vapour diffusion method, using 100 mM Tris (pH 8.5), 200 mM NaCl, and 24% PEG 4000 | Escherichia coli |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 3.2.2.8 | Q227A | the mutation causes an increase of kcat for uridine and inosine | Escherichia coli |
| 3.2.2.8 | Q227F | the mutation causes an increase of kcat for uridine and inosine | Escherichia coli |
| 3.2.2.8 | Q227Y | the mutation has a strong, enhancing effect on the hydrolysis of inosine, and the catalytic efficiency for the purinic substrate is increased by a factor of 7.6 | Escherichia coli |
| 3.2.2.8 | T223A | the mutation does not improve significantly the catalytic efficiency of YeiK toward inosine | Escherichia coli |
| 3.2.2.8 | T223F | the mutation does not improve significantly the catalytic efficiency of YeiK toward inosine | Escherichia coli |
| 3.2.2.8 | T223F/Q227Y | the mutant shows a 2fold increase in catalytic efficiency toward inosine | Escherichia coli |
| 3.2.2.8 | T223Y | the mutation does not affect the specificity of the enzyme toward inosine or uridine | Escherichia coli |
| 3.2.2.8 | T223Y/Q227Y | the mutant displays a catalytic efficiency toward inosine that is more than 50fold increased compared to that of wild type enzyme | Escherichia coli |
| 3.2.2.8 | T227A | the mutation does not improve significantly the catalytic efficiency of YeiK toward inosine | Escherichia coli |
| 3.2.2.8 | T227F | the mutation does not improve significantly the catalytic efficiency of YeiK toward inosine | Escherichia coli |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.2.2.8 | 0.12 | - |
uridine | wild type enzyme, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli |  |
| 3.2.2.8 | 0.31 | - |
uridine | mutant enzyme T223F, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 0.33 | - |
uridine | mutant enzyme T223A, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 0.57 | - |
uridine | mutant enzyme Q227Y, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 0.77 | - |
uridine | mutant enzyme Q227A, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 1.06 | - |
uridine | mutant enzyme T223Y/Q227Y, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 1.09 | - |
uridine | mutant enzyme T223F/Q227Y, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 1.13 | - |
uridine | mutant enzyme T223Y, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 1.16 | - |
Inosine | mutant enzyme T223Y, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 1.19 | - |
uridine | mutant enzyme Q227F, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 1.77 | - |
Inosine | mutant enzyme Q227F, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 1.93 | - |
Inosine | mutant enzyme T223Y/Q227Y, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 2.14 | - |
Inosine | mutant enzyme Q227Y, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 2.34 | - |
Inosine | wild type enzyme, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 3.29 | - |
Inosine | mutant enzyme T223A, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 4.31 | - |
Inosine | mutant enzyme Q227A, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 4.42 | - |
Inosine | mutant enzyme T223F/Q227Y, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 5.3 | - |
Inosine | mutant enzyme T223F, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 3.2.2.8 | Ca2+ | the substrate binds to the Ca2+-containing active site in the catalytic cavity at the C-terminal end of the core beta-sheet | Escherichia coli | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 3.2.2.8 | Escherichia coli | P33022 | - |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 3.2.2.8 | Ni-NTA column chromatography and MonoQ column chromatography | Escherichia coli |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.2.2.8 | inosine + H2O | poor substrate | Escherichia coli | hypoxanthine + D-ribose | - |
? | |
| 3.2.2.8 | uridine + H2O | - |
Escherichia coli | uracil + D-ribose | - |
? | |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 3.2.2.8 | CU-NH | - |
Escherichia coli |
| 3.2.2.8 | cytidine-uridine-preferring nucleoside hydrolase | - |
Escherichia coli |
| 3.2.2.8 | RihB | - |
Escherichia coli |
| 3.2.2.8 | YeiK | - |
Escherichia coli |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.2.2.8 | 0.035 | - |
Inosine | mutant enzyme T223Y, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 0.086 | - |
Inosine | wild type enzyme, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 0.109 | - |
Inosine | mutant enzyme T223F, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 0.125 | - |
Inosine | mutant enzyme Q227F, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 0.18 | - |
Inosine | mutant enzyme T223A, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 0.182 | - |
Inosine | mutant enzyme Q227A, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 0.382 | - |
Inosine | mutant enzyme T223F/Q227Y, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 0.593 | - |
Inosine | mutant enzyme Q227Y, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 3.62 | - |
Inosine | mutant enzyme T223Y/Q227Y, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 5.4 | - |
uridine | wild type enzyme, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 15.1 | - |
uridine | mutant enzyme Q227Y, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 18.8 | - |
uridine | mutant enzyme T223F, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 39.8 | - |
uridine | mutant enzyme T223A, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 44.3 | - |
uridine | mutant enzyme T223Y, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 46.9 | - |
uridine | mutant enzyme Q227A, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 52.5 | - |
uridine | mutant enzyme Q227F, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 59.1 | - |
uridine | mutant enzyme T223F/Q227Y, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
| 3.2.2.8 | 72.9 | - |
uridine | mutant enzyme T223Y/Q227Y, in 50 mM HEPES buffer (pH 7.3), at 37°C | Escherichia coli | |
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