Cloned (Comment) | Organism |
---|---|
pET-11a for expression in Escherichia coli BL21 (DE3) | Thermus thermophilus |
Crystallization (Comment) | Organism |
---|---|
crystallized at 18°C using the oil-microbatch method with PEG 4000 as a precipitant. A native data set is collected to 1.8 A resolution using synchrotron radiation. The crystal belongs to the monoclinic space group P2(1), with unit-cell parameters a = 58.83, b = 76.23, c = 103.86 A, beta = 91.3 | Thermus thermophilus |
crystals: monoclinic with dimensions of 0.13 * 0.13 * 0.07 mm, X-ray diffraction to 1.8 Å using synchrotron radiation at -173°C, belong to space group P2(1), asymmetric unit contains dimer with local pseudo-twofold symmetry, unit-cell parameters: a: 58.83, b: 76.23, c: 103.86, beta: 91.3°, oil-microbatch method: 1 week at 18°C, drop: 0.5 microlitre precipitant solution (27.5% (w/v) PEG 4000 in 100 mM HEPES-NaOH pH 7.5, 10 mM CaCl2) + 0.5 microlitre protein solution (21.7 mg/ml in 20 mM Tris-HCl pH8, 200 NaCl) | Thermus thermophilus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
45400 | - |
estimated according to amino acid composition | Thermus thermophilus |
86300 | - |
in the dimeric state of the purified protein | Thermus thermophilus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
pyrimidine nucleoside + phosphate | Thermus thermophilus | most pyrimidine nucleosides except for 4-amino-substituted species like deoxycytidine | pyrimidine base + alpha-D-ribose 1-phosphate | or alpha-D-deoxyribose 1-phosphate | r | |
pyrimidine nucleoside + phosphate | Thermus thermophilus HB8 / ATCC 27634 / DSM 579 | most pyrimidine nucleosides except for 4-amino-substituted species like deoxycytidine | pyrimidine base + alpha-D-ribose 1-phosphate | or alpha-D-deoxyribose 1-phosphate | r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermus thermophilus | - |
- |
- |
Thermus thermophilus | Q5SHF9 | HB8 | - |
Thermus thermophilus HB8 / ATCC 27634 / DSM 579 | - |
- |
- |
Thermus thermophilus HB8 / ATCC 27634 / DSM 579 | Q5SHF9 | HB8 | - |
Purification (Comment) | Organism |
---|---|
harvested with 5 mM 2-mercaptoethanol and 500 mM NaCl in Tris-HCl pH 8, extract desalted with HiPrep 2610, recombinant protein eluted with 0-300 mM NaCl from Super Q Toyopearl 650M column, eluted from Bio-Scale CHT-20-1 column with 10-100 mM NaCl in phosphate-NaOH pH 7, concentrated by ultracentrifugation with 10 kDa cut-off, eluted from HiLoad 16/60 Superdex 200 prep-grade column with 200 mM NaCl in Tris-HCl pH 8, concentrated by ultracentrifugation to 21.7 mg/ml, stored at -70 °C | Thermus thermophilus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
pyrimidine nucleoside + phosphate | most pyrimidine nucleosides except for 4-amino-substituted species like deoxycytidine | Thermus thermophilus | pyrimidine base + alpha-D-ribose 1-phosphate | or alpha-D-deoxyribose 1-phosphate | r | |
pyrimidine nucleoside + phosphate | most pyrimidine nucleosides except for 4-amino-substituted species like deoxycytidine | Thermus thermophilus HB8 / ATCC 27634 / DSM 579 | pyrimidine base + alpha-D-ribose 1-phosphate | or alpha-D-deoxyribose 1-phosphate | r | |
thymidine + phosphate | - |
Thermus thermophilus | thymine + alpha-D-deoxyribose 1-phosphate | - |
r | |
thymidine + phosphate | - |
Thermus thermophilus HB8 / ATCC 27634 / DSM 579 | thymine + alpha-D-deoxyribose 1-phosphate | - |
r |
Subunits | Comment | Organism |
---|---|---|
dimer | revealed by dynamic light-scattering (DynaPro MS/X) of the purified protein, 20 mg protein/ml, 18°C, 20 mM Tris-HCl pH 7.6, 200 mM sodium chloride, particle size distribution with a polydispersity of 18.7% | Thermus thermophilus |
Synonyms | Comment | Organism |
---|---|---|
PYNP | - |
Thermus thermophilus |
pyrimidine nucleoside phosphorylase | - |
Thermus thermophilus |
TTHA1771 | - |
Thermus thermophilus |