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Literature summary for 2.4.2.4 extracted from
- Structures of native human thymidine phosphorylase and in complex with 5-iodouracil (2009), Biochem. Biophys. Res. Commun., 386, 666-670.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression of wild-type and mutan enzymes in Escherichia coli strain BL21(DE3) | Homo sapiens |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| purified recombinant thymidine phosphorylase, free and in complex with 5-iodouracil, X-ray diffraction structure determination and analysis at 3.0 A and 2.5 A resolutions, respectively | Homo sapiens |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| H116F | site-directed mutagenesis, inactive mutant | Homo sapiens |
| H116K | site-directed mutagenesis, inactive mutant | Homo sapiens |
| I214A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
| K115A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
| K115E | site-directed mutagenesis, inactive mutant | Homo sapiens |
| R202E | site-directed mutagenesis, inactive mutant | Homo sapiens |
| R202S | site-directed mutagenesis, inactive mutant | Homo sapiens |
| S217G | site-directed mutagenesis, inactive mutant | Homo sapiens |
| Y199A | site-directed mutagenesis, inactive mutant | Homo sapiens |
| Y199F | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
| Y199L | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 5-Iodouracil | His116 directly interacts with the inhibitor via its NE2 group, enzyme binding structure, overview | Homo sapiens |  |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.35 | - |
thymidine | ph 5.5, 37°C, recombinant wild-type enzyme | Homo sapiens | |
| 0.42 | - |
thymidine | ph 5.5, 37°C, recombinant mutant K115A | Homo sapiens | |
| 0.74 | - |
thymidine | ph 5.5, 37°C, recombinant mutant I214A | Homo sapiens | |
| 2.1 | - |
thymidine | ph 5.5, 37°C, recombinant mutant Y199F | Homo sapiens | |
| 3.14 | - |
thymidine | ph 5.5, 37°C, recombinant mutant Y199L | Homo sapiens | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| thymidine + phosphate | Homo sapiens | - |
thymine + 2-deoxy-alpha-D-ribose 1-phosphate | - |
r | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Homo sapiens | P19971 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant wild-type and mutan enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration | Homo sapiens |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
- |
Homo sapiens |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| thymidine + phosphate | - |
Homo sapiens | thymine + 2-deoxy-alpha-D-ribose 1-phosphate | - |
r | |
| thymidine + phosphate | the enzyme catalyzes the reversible cleavage of the glycosidic bond of pyrimidine 2'-deoxynucleotides, most likely through SN2-like transition state involving nucleobase, 2'-deoxyribose and phosphate. The active site consists of residues His116, Ser117, Leu148, Arg202, Val208, Ile214, Lys221 and Val241 | Homo sapiens | thymine + 2-deoxy-alpha-D-ribose 1-phosphate | - |
r | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| More | the enzyme belongs to the pyrimidine nucleoside phosphorylase family | Homo sapiens |
| PDECGF | - |
Homo sapiens |
| platelet derived endothelial cell growth factor | - |
Homo sapiens |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Homo sapiens |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 3.5 | - |
thymidine | ph 5.5, 37°C, recombinant mutant Y199F | Homo sapiens | |
| 4.4 | - |
thymidine | ph 5.5, 37°C, recombinant mutant Y199L | Homo sapiens | |
| 5.2 | - |
thymidine | ph 5.5, 37°C, recombinant mutant I214A | Homo sapiens | |
| 6.8 | - |
thymidine | ph 5.5, 37°C, recombinant mutant K115A | Homo sapiens | |
| 8.2 | - |
thymidine | ph 5.5, 37°C, recombinant wild-type enzyme | Homo sapiens | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 5.5 | - |
thymine production, assay at | Homo sapiens |
Ki Value [mM]
| Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.48 | - |
5-Iodouracil | - |
Homo sapiens | |
General Information
| General Information | Comment | Organism |
|---|---|---|
| metabolism | the enzyme plays a key role in nucleoside metabolism | Homo sapiens |
| physiological function | the enzyme is the sole endothelial mitogenic and angiogenic factor, it plays a role in the stimulation of chemotaxis and thymidine incorporation into endothelial cells in vitro and angiogenesis in vivo | Homo sapiens |
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