Cloned (Comment) | Organism |
---|---|
from genomic DNA in pGEM-T-Eeasy, in pET15b for expression with hexa-His tag in Escherichia coli B21 Star (DE3) | Pyrococcus horikoshii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | no significant change in KM with temperature (40-70°C) for all substrates tested | Pyrococcus horikoshii | |
0.00326 | - |
guanine | +/-0.00029 mM, 70°C | Pyrococcus horikoshii | |
0.00388 | - |
hypoxanthine | +/-0.00033 mM, 40°C | Pyrococcus horikoshii | |
0.00483 | - |
hypoxanthine | +/-0.00049 mM, 60°C | Pyrococcus horikoshii | |
0.00485 | - |
guanine | +/-0.00056 mM, 50°C | Pyrococcus horikoshii | |
0.0061 | - |
hypoxanthine | +/-0.00143 mM, 70°C | Pyrococcus horikoshii | |
0.00702 | - |
hypoxanthine | +/-0.00104 mM, 50°C | Pyrococcus horikoshii | |
0.0282 | - |
5-phospho-alpha-D-ribose 1-diphosphate | +/-0.0015 mM, 60°C | Pyrococcus horikoshii | |
0.0369 | - |
5-phospho-alpha-D-ribose 1-diphosphate | +/-0.0024 mM, 70°C | Pyrococcus horikoshii | |
0.0412 | - |
5-phospho-alpha-D-ribose 1-diphosphate | +/-0.0016 mM, 50°C | Pyrococcus horikoshii | |
0.0428 | - |
5-phospho-alpha-D-ribose 1-diphosphate | +/-0.0009 mM, 40°C | Pyrococcus horikoshii | |
0.095 | - |
xanthine | +/-0.0084 mM, 70°C | Pyrococcus horikoshii | |
0.1012 | - |
xanthine | +/-0.0101 mM, 50°C | Pyrococcus horikoshii |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
103000 | - |
dynamic light scattering | Pyrococcus horikoshii |
103100 | - |
gel filtration chromatography | Pyrococcus horikoshii |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
guanine + 5-phospho-alpha-D-ribose 1-diphosphate | Pyrococcus horikoshii | - |
GMP + diphosphate | - |
? | |
hypoxanthine + 5-phospho-alpha-D-ribose 1-diphosphate | Pyrococcus horikoshii | - |
IMP + diphosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pyrococcus horikoshii | - |
hyperthermophile | - |
Purification (Comment) | Organism |
---|---|
thermal treatment (1 h, 70°C), immobilized metal affinity chromatography with Hi-Trap nickel chelating column (pH 7.5, elution with 200 mM imidazole), cleavage of His-tag with thrombin followed by second IMAC | Pyrococcus horikoshii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
guanine + 5-phospho-alpha-D-ribose 1-diphosphate | - |
Pyrococcus horikoshii | GMP + diphosphate | - |
? | |
guanine + 5-phospho-alpha-D-ribose 1-diphosphate | hypoxanthine and guanine preferred substrates | Pyrococcus horikoshii | GMP + diphosphate | - |
ir | |
hypoxanthine + 5-phospho-alpha-D-ribose 1-diphosphate | - |
Pyrococcus horikoshii | IMP + diphosphate | - |
? | |
hypoxanthine + 5-phospho-alpha-D-ribose 1-diphosphate | hypoxanthine and guanine preferred substrates, continuous spectrophotometric enzyme assay, pH 7.5, 12 mM MgCl2, 40-70°C, 30 sec | Pyrococcus horikoshii | IMP + diphosphate | Kd(diphosphate): 4.7 +/-0.1 mM, revealed by quenching of intrinsic fluorescence of the enzyme upon diphosphate-binding | ir | |
additional information | no formation of PRPP and 6-oxopurine by pyrophosphorylysis due to disability of binding to nucleotides IMP, GMP, XMP at 40-70°C and substrate concentrations of up to 0.5 mM | Pyrococcus horikoshii | ? | - |
? | |
xanthine + 5-phospho-alpha-D-ribose 1-diphosphate | low catalytic efficiency (kcat/KM), xanthine is not preferred but a usable substrate | Pyrococcus horikoshii | XMP + diphosphate | - |
ir |
Subunits | Comment | Organism |
---|---|---|
homohexamer | dynamic light scattering (pH 7.5, 25 °C) and gel filtration chromatography (HR 10/30 Superdex 200, FPLC, pH 7.5) | Pyrococcus horikoshii |
Synonyms | Comment | Organism |
---|---|---|
6-oxopurine phosphoribosyltransferase | - |
Pyrococcus horikoshii |
HGXPRT | - |
Pyrococcus horikoshii |
HPRT | - |
Pyrococcus horikoshii |
hypoxanthine-guanine phosphoribosyltransferase | - |
Pyrococcus horikoshii |
hypoxanthine-guanine-xanthine phosphoribosyltransferase | - |
Pyrococcus horikoshii |
PhHGXPRT | Pyrococcus horikoshii HGXPRT | Pyrococcus horikoshii |
PhPRT | - |
Pyrococcus horikoshii |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | increasing kcat with temperature (40-70°C) for all substrates tested | Pyrococcus horikoshii | |
additional information | - |
additional information | kcat/KM for 5-phospho-alpha-D-ribose 1-diphosphate = 0.00093 mM(-1)*min(-1) at 40°C, 0.00069 mM(-1)*min(-1) at 50°C, 0.00357 mM(-1)*min(-1) at 60°C, 0.00496 mM(-1)*min(-1) at 70°C | Pyrococcus horikoshii | |
additional information | - |
additional information | kcat/KM for guanine = 0.00262 mM(-1)*min(-1) at 50°C, 0.02631 mM(-1)*min(-1) at 70°C | Pyrococcus horikoshii | |
additional information | - |
additional information | kcat/KM for hypoxanthine = 0.00579 mM(-1)*min(-1) at 40°C, 0.00708 mM(-1)*min(-1) at 50°C, 0.01719 mM(-1)*min(-1) at 60°C, 0.03107 mM(-1)*min(-1) at 70°C | Pyrococcus horikoshii | |
additional information | - |
additional information | kcat/KM for hypoxanthine similar to kcat/KM for guanine, kcat/KM for xanthine approximately 20 times lower | Pyrococcus horikoshii | |
additional information | - |
additional information | kcat/KM for xanthine = 0.00040 mM(-1)*min(-1) at 50°C, 0.00139 mM(-1)*min(-1) at 70°C | Pyrococcus horikoshii | |
0.21 | - |
guanine | +/-0.01, 50°C | Pyrococcus horikoshii | |
0.38 | - |
hypoxanthine | +/-0.01, 40°C | Pyrococcus horikoshii | |
0.48 | - |
5-phospho-alpha-D-ribose 1-diphosphate | +/-0.02, 50°C | Pyrococcus horikoshii | |
0.66 | - |
5-phospho-alpha-D-ribose 1-diphosphate | +/-0.03, 40°C | Pyrococcus horikoshii | |
0.68 | - |
xanthine | +/-0.04, 50°C | Pyrococcus horikoshii | |
0.83 | - |
hypoxanthine | +/-0.04, 50°C | Pyrococcus horikoshii | |
1.38 | - |
hypoxanthine | +/-0.03, 60°C | Pyrococcus horikoshii | |
1.43 | - |
guanine | +/-0.02, 70°C | Pyrococcus horikoshii | |
1.68 | - |
5-phospho-alpha-D-ribose 1-diphosphate | +/-0.06, 60°C | Pyrococcus horikoshii | |
2.2 | - |
xanthine | +/-0.08, 70°C | Pyrococcus horikoshii | |
3.05 | - |
5-phospho-alpha-D-ribose 1-diphosphate | +/-0.07, 70°C | Pyrococcus horikoshii | |
3.16 | - |
hypoxanthine | +/-0.08, 70°C | Pyrococcus horikoshii |