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Literature summary extracted from

  • Park, J.I.; Cao, L.; Platt, V.M.; Huang, Z.; Stull, R.A.; Dy, E.E.; Sperinde, J.J.; Yokoyama, J.S.; Szoka, F.C.
    Antitumor therapy mediated by 5-fluorocytosine and a recombinant fusion protein containing TSG-6 hyaluronan binding domain and yeast cytosine deaminase (2009), Mol. Pharm., 6, 801-812.
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
3.5.4.1 medicine the hyaluronan binding domain of tumor necrosis factor alpha-stimulated gene-6-cytosine deaminase fusion protein is an alternative to antibody-directed prodrug enzyme therapy approaches for cancer treatment Saccharomyces cerevisiae

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.4.1 expressed in Escherichia coli BL21-Codon Plus (DE3)-RIPL cells Saccharomyces cerevisiae

Protein Variants

EC Number Protein Variants Comment Organism
3.5.4.1 E64A the mutant lacks enzyme activity Saccharomyces cerevisiae

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.4.1 0.33
-
 5-fluorocytosine purified hyaluronan binding domain of TSG-6-cytosine deaminase fusion protein, in PBS buffer, pH not specified in the publication, at 37°C Saccharomyces cerevisiae
3.5.4.1 0.51
-
 5-fluorocytosine recombinant cytosine deaminase, in PBS buffer, pH not specified in the publication, at 37°C Saccharomyces cerevisiae

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.5.4.1 19000
-
 2 * 19000, theoretical value Saccharomyces cerevisiae
3.5.4.1 29000
-
 gel filtration Saccharomyces cerevisiae

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.5.4.1 cytosine + H2O Saccharomyces cerevisiae
-
 uracil + NH3
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.5.4.1 Saccharomyces cerevisiae
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.4.1 Ni2+-NTA column chromatography Saccharomyces cerevisiae

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.4.1 5-fluorocytosine + H2O
-
 Saccharomyces cerevisiae 5-fluorouracil + NH3
-
 ?
3.5.4.1 cytosine + H2O
-
 Saccharomyces cerevisiae uracil + NH3
-
 ?

Subunits

EC Number Subunits Comment Organism
3.5.4.1 homodimer 2 * 19000, theoretical value Saccharomyces cerevisiae

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.5.4.1 37
-
 the hyaluronan binding domain of tumor necrosis factor alpha-stimulated gene-6-cytosine deaminase fusion protein retains 20% of its initial enzyme activity after 24 h, 12% after 48 h, and 6% after 124 h at 37°C. In contrast, the activity of cytosine deaminase decreases rapidly at 37°C, and retains only about 5% of initial activity after 24 h Saccharomyces cerevisiae
3.5.4.1 42
-
 the melting temperature of cytosine deaminase is at 42°C Saccharomyces cerevisiae