EC Number | Cloned (Comment) | Organism |
---|---|---|
3.2.2.1 | recombinant expression of His-tagged wild-type and mutant enzymes | Physcomitrium patens |
3.2.2.1 | the gene is located on chromosome 2, expression of His-tagged wild-type and mutant enzymes | Zea mays |
3.2.2.3 | isozyme PpNRH2, phylogenetic analysis, recombinant expression in Escherichia coli in inclusion bodies | Physcomitrium patens |
3.2.2.3 | phylogenetic analysis | Zea mays |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
3.2.2.1 | recombinant His-tagged wild-type enzyme, hanging drop avpour diffusion method, 30-35 mg/ml protein is mixed with an equal volume of a precipitant solution containing 0.1 M HEPES, pH 7.5, 100 mM sodium acetate, 10% w/v PEG 4000, and 10% ethylene glycol, X-ray diffraction structure determination and analysis at 3.35 A resolution, molecular replacement | Physcomitrium patens |
3.2.2.1 | recombinant His-tagged wild-type enzyme, hanging drop avpour diffusion method, 35 mg/ml protein is mixed with an equal volume of reservori solution containing 50 mM Tris-HCl, pH 8.0, 150 mM NaCl, and 20% w/v PEG 2000 monomethyl ether, X-ray diffraction structure determination and analysis at 2.49 A resolution, molecular replacement | Zea mays |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
3.2.2.1 | D250A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Physcomitrium patens |
3.2.2.1 | D252A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Physcomitrium patens |
3.2.2.1 | D25A | site-directed mutagenesis, inactive mutant | Physcomitrium patens |
3.2.2.1 | D8A | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Zea mays |
3.2.2.1 | E247A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Physcomitrium patens |
3.2.2.1 | H245A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Physcomitrium patens |
3.2.2.1 | H99A | site-directed mutagenesis, inactive mutant | Physcomitrium patens |
3.2.2.1 | additional information | generation of a functional gene knockout of PpNRH1 using a gene-replacement vector. Phenotype and Growth of enzyme knockout mutant in medium with nucleosides as the sole nitrogen source, overview | Physcomitrium patens |
3.2.2.1 | Y241A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Physcomitrium patens |
3.2.2.1 | Y244A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Physcomitrium patens |
3.2.2.1 | Y249A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Physcomitrium patens |
3.2.2.1 | Y255A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Physcomitrium patens |
3.2.2.3 | additional information | functional knockout of the mutant, phenotypic analysis of wild-type and mutant enzymes, overview | Physcomitrium patens |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.2.2.1 | additional information | - |
additional information | substrate specificity and kinetics, overview | Zea mays | |
3.2.2.1 | 0.078 | - |
Inosine | pH 7.5, 30°C, wild-type enzyme | Physcomitrium patens | |
3.2.2.1 | 0.094 | - |
Inosine | pH 7.5, 30°C, mutant E247A | Physcomitrium patens | |
3.2.2.1 | 0.116 | - |
Xanthosine | pH 7.5, 30°C, wild-type enzyme | Physcomitrium patens | |
3.2.2.1 | 0.134 | - |
Inosine | pH 7.5, 30°C, mutant D250A | Physcomitrium patens | |
3.2.2.1 | 0.14 | - |
Xanthosine | pH 7.5, 30°C, mutant E247A | Physcomitrium patens | |
3.2.2.1 | 0.171 | - |
Xanthosine | pH 7.5, 30°C, mutant D250A | Physcomitrium patens | |
3.2.2.1 | 0.201 | - |
Inosine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.1 | 0.206 | - |
Xanthosine | pH 7.5, 30°C, mutant Y244A | Physcomitrium patens | |
3.2.2.1 | 0.232 | - |
Xanthosine | pH 7.5, 30°C, mutant D252A | Physcomitrium patens | |
3.2.2.1 | 0.301 | - |
Inosine | pH 7.5, 30°C, mutant D252A | Physcomitrium patens | |
3.2.2.1 | 0.356 | - |
Xanthosine | pH 7.5, 30°C, mutant H245A | Physcomitrium patens | |
3.2.2.1 | 0.367 | - |
Xanthosine | pH 7.5, 30°C, mutant Y255A | Physcomitrium patens | |
3.2.2.1 | 0.396 | - |
Xanthosine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.1 | 0.487 | - |
Xanthosine | pH 7.5, 30°C, mutant Y241A | Physcomitrium patens | |
3.2.2.1 | 0.596 | - |
Xanthosine | pH 7.5, 30°C, mutant Y249A | Physcomitrium patens | |
3.2.2.1 | 0.63 | - |
Inosine | pH 7.5, 30°C, mutant Y241A | Physcomitrium patens | |
3.2.2.1 | 0.76 | - |
Inosine | pH 7.5, 30°C, mutant Y255A | Physcomitrium patens | |
3.2.2.1 | 1.09 | - |
Inosine | pH 7.5, 30°C, mutant H245A | Physcomitrium patens | |
3.2.2.1 | 1.208 | - |
Inosine | pH 7.5, 30°C, mutant Y244A | Physcomitrium patens | |
3.2.2.1 | 1390 | - |
Inosine | pH 7.5, 30°C, mutant Y249A | Physcomitrium patens | |
3.2.2.3 | additional information | - |
additional information | substrate specificity and kinetics, overview | Zea mays | |
3.2.2.3 | 0.06 | - |
adenosine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.3 | 0.109 | - |
Xanthosine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.3 | 0.111 | - |
adenosine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.3 | 0.178 | - |
Xanthosine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.3 | 0.468 | - |
uridine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.3 | 0.512 | - |
uridine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.3 | 0.713 | - |
Inosine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.3 | 1.013 | - |
Inosine | pH 7.5, 30°C, recombinant enzyme | Zea mays |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.2.2.3 | uridine + H2O | Zea mays | - |
D-ribose + uracil | - |
? | |
3.2.2.3 | uridine + H2O | Physcomitrium patens | - |
D-ribose + uracil | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
3.2.2.1 | Physcomitrium patens | A9TXA6 | - |
- |
3.2.2.1 | Physcomitrium patens Gransden 2004 | A9TXA6 | - |
- |
3.2.2.1 | Zea mays | B6T563 | - |
- |
3.2.2.3 | Physcomitrium patens | - |
- |
- |
3.2.2.3 | Zea mays | - |
- |
- |
3.2.2.3 | Zea mays | H9D3U7 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
3.2.2.1 | recombinant His-tagged wild-type and mutant enzymes by cobalt affinity chromatography and gel filtration | Physcomitrium patens |
3.2.2.1 | recombinant His-tagged wild-type and mutant enzymes by cobalt affinity chromatography and gel filtration | Zea mays |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.2.2.1 | inosine + H2O | best substrate | Zea mays | D-ribose + hypoxanthine | - |
? | |
3.2.2.1 | inosine + H2O | 87% of the activity with xanthosine | Physcomitrium patens | D-ribose + hypoxanthine | - |
? | |
3.2.2.1 | inosine + H2O | 87% of the activity with xanthosine | Physcomitrium patens Gransden 2004 | D-ribose + hypoxanthine | - |
? | |
3.2.2.1 | additional information | poor activity with adenosine, uridine, and guanosine, no activity with cytokinin riboside, cytidine, and trans-zeatin riboside. Substrate docking into the active site, overview | Physcomitrium patens | ? | - |
? | |
3.2.2.1 | additional information | poor activity with adenosine, uridine, guanosine, trans-zeatin riboside, and cytokinin riboside, almost no activity with cytidine. Substrate docking into the active site, overview | Zea mays | ? | - |
? | |
3.2.2.1 | additional information | poor activity with adenosine, uridine, and guanosine, no activity with cytokinin riboside, cytidine, and trans-zeatin riboside. Substrate docking into the active site, overview | Physcomitrium patens Gransden 2004 | ? | - |
? | |
3.2.2.1 | xanthosine + H2O | best substrate | Physcomitrium patens | D-ribose + xanthine | - |
? | |
3.2.2.1 | xanthosine + H2O | 70% of the activity with inosine | Zea mays | D-ribose + xanthine | - |
? | |
3.2.2.1 | xanthosine + H2O | best substrate | Physcomitrium patens Gransden 2004 | D-ribose + xanthine | - |
? | |
3.2.2.3 | adenosine + H2O | 15% of the activity with uridine | Zea mays | D-ribose + adenine | - |
? | |
3.2.2.3 | adenosine + H2O | 3.5% of the activity with uridine | Zea mays | D-ribose + adenine | - |
? | |
3.2.2.3 | cytidine + H2O | 3.9% of the activity with uridine | Physcomitrium patens | D-ribose + cytosine | - |
? | |
3.2.2.3 | cytidine + H2O | 5.0% of the activity with uridine | Zea mays | D-ribose + cytosine | - |
? | |
3.2.2.3 | inosine + H2O | 22% of the activity with uridine | Physcomitrium patens | D-ribose + hypoxanthine | - |
? | |
3.2.2.3 | inosine + H2O | 9.3% of the activity with uridine | Zea mays | D-ribose + hypoxanthine | - |
? | |
3.2.2.3 | inosine + H2O | 9.5% of the activity with uridine | Zea mays | D-ribose + hypoxanthine | - |
? | |
3.2.2.3 | additional information | adenosine is a poor substrates, no activity with guanosine, trans-zeatin riboside, and cytokinin riboside, substrate specificity, overview | Physcomitrium patens | ? | - |
? | |
3.2.2.3 | additional information | no or almost no activity with guanosine, trans-zeatin riboside, and cytokinin riboside, substrate specificity, overview | Zea mays | ? | - |
? | |
3.2.2.3 | additional information | poor activity with cytidine, guanosine, trans-zeatin riboside, and cytokinin riboside, substrate specificity, overview | Zea mays | ? | - |
? | |
3.2.2.3 | uridine + H2O | - |
Zea mays | D-ribose + uracil | - |
? | |
3.2.2.3 | uridine + H2O | - |
Physcomitrium patens | D-ribose + uracil | - |
? | |
3.2.2.3 | uridine + H2O | best substrate | Zea mays | D-ribose + uracil | - |
? | |
3.2.2.3 | uridine + H2O | best substrate | Physcomitrium patens | D-ribose + uracil | - |
? | |
3.2.2.3 | xanthosine + H2O | 34% of the activity with uridine | Zea mays | D-ribose + xanthine | - |
? | |
3.2.2.3 | xanthosine + H2O | 53% of the activity with uridine | Zea mays | D-ribose + xanthine | - |
? | |
3.2.2.3 | xanthosine + H2O | 61% of the activity with uridine | Physcomitrium patens | D-ribose + xanthine | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
3.2.2.1 | PpNRH1 | - |
Physcomitrium patens |
3.2.2.1 | purine NRH | - |
Physcomitrium patens |
3.2.2.1 | purine NRH | - |
Zea mays |
3.2.2.1 | purine nucleoside N-ribohydrolase | - |
Physcomitrium patens |
3.2.2.1 | purine nucleoside N-ribohydrolase | - |
Zea mays |
3.2.2.1 | ZmNRH3 | - |
Zea mays |
3.2.2.3 | PpNRH2 | - |
Physcomitrium patens |
3.2.2.3 | ZmNRH2a | - |
Zea mays |
3.2.2.3 | ZmNRH2b | - |
Zea mays |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
3.2.2.1 | 30 | - |
assay at | Physcomitrium patens |
3.2.2.1 | 30 | - |
assay at | Zea mays |
3.2.2.3 | 30 | - |
assay at | Zea mays |
3.2.2.3 | 30 | - |
assay at | Physcomitrium patens |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.2.2.1 | 0.06 | - |
Inosine | pH 7.5, 30°C, mutant Y241A | Physcomitrium patens | |
3.2.2.1 | 0.19 | - |
Xanthosine | pH 7.5, 30°C, mutant Y241A | Physcomitrium patens | |
3.2.2.1 | 0.43 | - |
Xanthosine | pH 7.5, 30°C, mutant Y255A | Physcomitrium patens | |
3.2.2.1 | 0.47 | - |
Inosine | pH 7.5, 30°C, mutant Y255A | Physcomitrium patens | |
3.2.2.1 | 0.58 | - |
Xanthosine | pH 7.5, 30°C, mutant Y244A | Physcomitrium patens | |
3.2.2.1 | 0.7 | - |
Inosine | pH 7.5, 30°C, mutant Y249A | Physcomitrium patens | |
3.2.2.1 | 1.1 | - |
Inosine | pH 7.5, 30°C, mutant H245A | Physcomitrium patens | |
3.2.2.1 | 1.1 | - |
Xanthosine | pH 7.5, 30°C, mutant Y249A | Physcomitrium patens | |
3.2.2.1 | 1.3 | - |
Xanthosine | pH 7.5, 30°C, mutant H245A | Physcomitrium patens | |
3.2.2.1 | 1.4 | - |
Inosine | pH 7.5, 30°C, mutant Y244A | Physcomitrium patens | |
3.2.2.1 | 2.5 | - |
Xanthosine | pH 7.5, 30°C, mutant D252A | Physcomitrium patens | |
3.2.2.1 | 3 | - |
Inosine | pH 7.5, 30°C, mutant D252A | Physcomitrium patens | |
3.2.2.1 | 5.7 | - |
Inosine | pH 7.5, 30°C, mutant E247A | Physcomitrium patens | |
3.2.2.1 | 6 | - |
Inosine | pH 7.5, 30°C, wild-type enzyme | Physcomitrium patens | |
3.2.2.1 | 6.2 | - |
Xanthosine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.1 | 7 | - |
Xanthosine | pH 7.5, 30°C, wild-type enzyme | Physcomitrium patens | |
3.2.2.1 | 7.2 | - |
Inosine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.1 | 7.2 | - |
Inosine | pH 7.5, 30°C, mutant D250A | Physcomitrium patens | |
3.2.2.1 | 7.2 | - |
Xanthosine | pH 7.5, 30°C, mutant E247A | Physcomitrium patens | |
3.2.2.1 | 8.4 | - |
Xanthosine | pH 7.5, 30°C, mutant D250A | Physcomitrium patens | |
3.2.2.3 | 0.18 | - |
adenosine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.3 | 0.4 | - |
adenosine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.3 | 0.61 | - |
Inosine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.3 | 1.3 | - |
Xanthosine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.3 | 3 | - |
uridine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.3 | 4.1 | - |
Inosine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.3 | 4.6 | - |
Xanthosine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.3 | 23.8 | - |
uridine | pH 7.5, 30°C, recombinant enzyme | Zea mays |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
3.2.2.1 | 7.5 | - |
assay at | Physcomitrium patens |
3.2.2.1 | 7.5 | - |
assay at | Zea mays |
3.2.2.3 | 7.5 | - |
assay at | Zea mays |
3.2.2.3 | 7.5 | - |
assay at | Physcomitrium patens |
EC Number | General Information | Comment | Organism |
---|---|---|---|
3.2.2.1 | malfunction | changes in the levels of purine, pyrimidine, and cytokinin metabolites in knockout mutants, phenotypes, overview | Physcomitrium patens |
3.2.2.1 | additional information | all plant nucleoside N-ribohydrolases exhibit a conserved sequence motif, DTDPGIDD, at the N-terminus, and the second Asp of the DTDPGIDD conserved motif functions as the active site base | Zea mays |
3.2.2.1 | additional information | the presence of a tyrosine at position 249 (PpNRH1 numbering) confers high hydrolase activity for purine ribosides. All plant nucleoside N-ribohydrolases exhibit a conserved sequence motif, DTDPGIDD, at the N-terminus, and the second Asp of the DTDPGIDD conserved motif functions as the active site base | Physcomitrium patens |
3.2.2.3 | malfunction | changes in the levels of purine, pyrimidine, and cytokinin metabolites in knockout mutants, phenotypes, overview | Physcomitrium patens |
3.2.2.3 | additional information | the presence of a tyrosine at position 249 (PpNRH1 numbering) confers high hydrolase activity for purine ribosides | Zea mays |
3.2.2.3 | additional information | the presence of a tyrosine at position 249 (PpNRH1 numbering) confers high hydrolase activity for purine ribosides | Physcomitrium patens |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.2.2.1 | 0.1 | - |
Inosine | pH 7.5, 30°C, mutant Y241A | Physcomitrium patens | |
3.2.2.1 | 0.4 | - |
Xanthosine | pH 7.5, 30°C, mutant Y241A | Physcomitrium patens | |
3.2.2.1 | 0.51 | - |
Inosine | pH 7.5, 30°C, mutant Y249A | Physcomitrium patens | |
3.2.2.1 | 0.62 | - |
Inosine | pH 7.5, 30°C, mutant Y255A | Physcomitrium patens | |
3.2.2.1 | 1 | - |
Inosine | pH 7.5, 30°C, mutant H245A | Physcomitrium patens | |
3.2.2.1 | 1.2 | - |
Inosine | pH 7.5, 30°C, mutant Y244A | Physcomitrium patens | |
3.2.2.1 | 1.2 | - |
Xanthosine | pH 7.5, 30°C, mutant Y255A | Physcomitrium patens | |
3.2.2.1 | 1.8 | - |
Xanthosine | pH 7.5, 30°C, mutant Y249A | Physcomitrium patens | |
3.2.2.1 | 2.8 | - |
Xanthosine | pH 7.5, 30°C, mutant Y244A | Physcomitrium patens | |
3.2.2.1 | 3.6 | - |
Xanthosine | pH 7.5, 30°C, mutant H245A | Physcomitrium patens | |
3.2.2.1 | 10 | - |
Inosine | pH 7.5, 30°C, mutant D252A | Physcomitrium patens | |
3.2.2.1 | 11 | - |
Xanthosine | pH 7.5, 30°C, mutant D252A | Physcomitrium patens | |
3.2.2.1 | 16 | - |
Xanthosine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.1 | 36 | - |
Inosine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.1 | 49 | - |
Xanthosine | pH 7.5, 30°C, mutant D250A | Physcomitrium patens | |
3.2.2.1 | 51 | - |
Xanthosine | pH 7.5, 30°C, mutant E247A | Physcomitrium patens | |
3.2.2.1 | 53 | - |
Inosine | pH 7.5, 30°C, mutant D250A | Physcomitrium patens | |
3.2.2.1 | 61 | - |
Xanthosine | pH 7.5, 30°C, wild-type enzyme | Physcomitrium patens | |
3.2.2.1 | 61 | - |
Inosine | pH 7.5, 30°C, mutant E247A | Physcomitrium patens | |
3.2.2.1 | 77 | - |
Inosine | pH 7.5, 30°C, wild-type enzyme | Physcomitrium patens | |
3.2.2.3 | 0.6 | - |
Inosine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.3 | 3 | - |
adenosine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.3 | 3.5 | - |
adenosine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.3 | 5.7 | - |
Inosine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.3 | 6.4 | - |
uridine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.3 | 7.5 | - |
Xanthosine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.3 | 42 | - |
Xanthosine | pH 7.5, 30°C, recombinant enzyme | Zea mays | |
3.2.2.3 | 46 | - |
uridine | pH 7.5, 30°C, recombinant enzyme | Zea mays |