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Literature summary extracted from

  • Shimizu, K.; Kunishima, N.
    Purification, crystallization and preliminary X-ray diffraction study on pyrimidine nucleoside phosphorylase TTHA1771 from Thermus thermophilus HB8 (2007), Acta Crystallogr. Sect. F, 63, 308-310.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.2.2 pET-11a for expression in Escherichia coli BL21 (DE3) Thermus thermophilus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.4.2.2 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
2.4.2.2 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]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.4.2.2 45400
-
 estimated according to amino acid composition Thermus thermophilus
2.4.2.2 86300
-
 in the dimeric state of the purified protein Thermus thermophilus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.2.2 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
2.4.2.2 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

EC Number Organism UniProt Comment Textmining
2.4.2.2 Thermus thermophilus
-
-
-
2.4.2.2 Thermus thermophilus Q5SHF9 HB8
-
2.4.2.2 Thermus thermophilus HB8 / ATCC 27634 / DSM 579
-
-
-
2.4.2.2 Thermus thermophilus HB8 / ATCC 27634 / DSM 579 Q5SHF9 HB8
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.2.2 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 and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.2.2 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
2.4.2.2 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
2.4.2.2 thymidine + phosphate
-
 Thermus thermophilus thymine + alpha-D-deoxyribose 1-phosphate
-
 r
2.4.2.2 thymidine + phosphate
-
 Thermus thermophilus HB8 / ATCC 27634 / DSM 579 thymine + alpha-D-deoxyribose 1-phosphate
-
 r

Subunits

EC Number Subunits Comment Organism
2.4.2.2 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

EC Number Synonyms Comment Organism
2.4.2.2 PYNP
-
 Thermus thermophilus
2.4.2.2 pyrimidine nucleoside phosphorylase
-
 Thermus thermophilus
2.4.2.2 TTHA1771
-
 Thermus thermophilus