Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutan enzymes in Escherichia coli strain BL21(DE3) | Homo sapiens |
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 | 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 | 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 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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P19971 | - |
- |
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 Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Homo sapiens |
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 | 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 Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Homo sapiens |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5.5 | - |
thymine production, assay at | Homo sapiens |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.48 | - |
5-Iodouracil | - |
Homo sapiens |
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 |