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Literature summary extracted from

  • Giabbai, B.; Degano, M.
    Cloning, purification, crystallization and X-ray analysis of the Escherichia coli pyrimidine nucleoside hydrolase YeiK (2004), Acta Crystallogr. Sect. D, 60, 524-527.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.2.8 gene yeiK, subcloning in strain DH5alpha, expression of the His6-tagged enzyme in strain BL21(DE3) Escherichia coli K-12

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.2.2.8 purified detagged recombinant enzyme, hanging drop vapour diffusion method, 10 mg/ml protein in 20 mM HEPES, pH 7.4, 150 mM NaCl, is mixed with precipitant solution, 25°C, equilibration versus reservoir solution containing 100 mM Tris-HCl, pH 8.5, 200 mM NaCl, 25% w/v PEG 3350, cryoprotection by 25% glycerol, X-ray diffraction structure determination and analysis at 1.7 A resolution Escherichia coli K-12

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.2.8 36000
-
 4 * 36000, detagged recombinant enzyme, SDS-PAGE Escherichia coli K-12
3.2.2.8 143000
-
 detagged recombinant enzyme, gel filtration Escherichia coli K-12

Organism

EC Number Organism UniProt Comment Textmining
3.2.2.8 Escherichia coli K-12
-
 gene yeiK
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.2.8 recombinant His6-tagged enzyme from strain BL21(DE3) by nickel chelate affinity chromatography and gel filtration, the His-tag is cleaved off by thrombin Escherichia coli K-12

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.2.8 5-fluorouridine + H2O detagged recombinant enzyme Escherichia coli K-12 5-fluorouracil + D-ribose
-
 ?
3.2.2.8 cytidine + H2O detagged recombinant enzyme Escherichia coli K-12 cytosine + D-ribose
-
 ?
3.2.2.8 additional information detagged recombinant enzyme: substrate specificity, no activity with purine nucleosides, and with 2'-, 3'-, and 5'-deoxynucleosides Escherichia coli K-12 ?
-
 ?
3.2.2.8 uridine + H2O detagged recombinant enzyme Escherichia coli K-12 uracil + D-ribose
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.2.8 More structure analysis by dynamic light scattering Escherichia coli K-12
3.2.2.8 tetramer 4 * 36000, detagged recombinant enzyme, SDS-PAGE Escherichia coli K-12

Synonyms

EC Number Synonyms Comment Organism
3.2.2.8 More the enzyme is a member of the nucleoside hydrolase enzyme family Escherichia coli K-12
3.2.2.8 pyrimidine nucleoside hydrolase
-
 Escherichia coli K-12
3.2.2.8 pyrimidine-specific nucleoside hydrolase
-
 Escherichia coli K-12
3.2.2.8 YeiK
-
 Escherichia coli K-12