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Literature summary extracted from

  • Helt, S.S.; Thymark, M.; Harris, P.; Aagaard, C.; Dietrich, J.; Larsen, S.; Willemoes, M.
    Mechanism of dTTP inhibition of the bifunctional dCTP deaminase:dUTPase encoded by Mycobacterium tuberculosis (2008), J. Mol. Biol., 376, 554-569.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.6.1.23 enzyme expression in Escherichia coli strain BL21(DE3) Mycobacterium tuberculosis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.6.1.23 purified recombinant bifunctional dCTP deaminase:dUTPase in complex with inhibitor thymidine triphosphate, hanging drop vapor diffusion method, 15°C, 0.004 ml of enzyme solution containing 1.8 mg/ml enzyme, 20 mM MgCl2, 5 mM dTTP, 50 mM HEPES, pH 6.8, is mixed with 0.002 ml of reservoir solution, cotaining 45% PEG 400, 200 mM MgCl2 and 100 mM HEPES, pH 7.5, and equilibrated over 1 ml of reservoir solution, 1 day, for the free enzyme crystallization is used: 1.9 mg/ml enzyme and 50 mM HEPES, pH 6.8, mixed with 0.002 ml of reservoir solution, 20% PEG 8000, 50 mM MgCl2 and 100 mM HEPES, pH 7.5, and equilibrated over 1 ml of reservoir solution, for 1 day to 6 weeks, X-ray diffraction structure determination and analysis at 2.0-2.5 A resolution Mycobacterium tuberculosis

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.4.13 dTTP the inhibitor has a regukatory function for the bifunctional enzyme, mechanism of regulation by conformational changes, overview Mycobacterium tuberculosis
3.6.1.23 thymidine triphosphate binding of thymidine triphosphate leads to disordered C-terminal arranged as a lid covering the active site, and the enzyme adapts an inactive conformation as a result of structural changes in the active site, mechanism, overview Mycobacterium tuberculosis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.4.13 additional information
-
 additional information steady-state kinetic analysis Mycobacterium tuberculosis
3.6.1.23 additional information
-
 additional information steady-state kinetic analysis of the dual activities of the bifunctional enzyme, overview Mycobacterium tuberculosis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.6.1.23 25000
-
 x * 25000, recombinant enzyme, SDS-PAGE Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.5.4.13 dCTP + H2O Mycobacterium tuberculosis
-
 dUTP + NH3
-
 ?
3.5.4.13 dCTP + H2O Mycobacterium tuberculosis H37Rv
-
 dUTP + NH3
-
 ?
3.6.1.23 dUTP + H2O Mycobacterium tuberculosis
-
 dUMP + diphosphate
-
 ?
3.6.1.23 dUTP + H2O Mycobacterium tuberculosis H37Rv
-
 dUMP + diphosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.5.4.13 Mycobacterium tuberculosis P9WP17
-
-
3.5.4.13 Mycobacterium tuberculosis H37Rv P9WP17
-
-
3.6.1.23 Mycobacterium tuberculosis P9WNS5
-
-
3.6.1.23 Mycobacterium tuberculosis H37Rv P9WNS5
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.6.1.23 recombinant enzyme from Escherichia coli strain BL21(DE3) by streptomycin precipitation, anion exchange chromatography, dialysis, and ammonium sulfate fractionation to homogeneity Mycobacterium tuberculosis

Reaction

EC Number Reaction Comment Organism Reaction ID
3.5.4.13 dCTP + H2O = dUTP + NH3 the mechanism for the deamination of dCTP involves the four residues Ser102, Arg106, Ala115 and Glu129, structure-function relationship of the bifunctional enzyme, overview Mycobacterium tuberculosis
a
3.6.1.23 dUTP + H2O = dUMP + diphosphate catalytic mechanism Mycobacterium tuberculosis
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.4.13 dCTP + H2O
-
 Mycobacterium tuberculosis dUTP + NH3
-
 ?
3.5.4.13 dCTP + H2O
-
 Mycobacterium tuberculosis H37Rv dUTP + NH3
-
 ?
3.5.4.13 additional information the enzyme is a bifunctional dCTP deaminase:dUTPase Mycobacterium tuberculosis ?
-
 ?
3.5.4.13 additional information the enzyme is a bifunctional dCTP deaminase:dUTPase Mycobacterium tuberculosis H37Rv ?
-
 ?
3.6.1.23 dUTP + H2O
-
 Mycobacterium tuberculosis dUMP + diphosphate
-
 ?
3.6.1.23 dUTP + H2O
-
 Mycobacterium tuberculosis H37Rv dUMP + diphosphate
-
 ?
3.6.1.23 additional information the bifunctional dCTP deaminase:dUTPase shows very similar affinities for dCTP and deoxyuridine triphosphate as substrates, overview Mycobacterium tuberculosis ?
-
 ?
3.6.1.23 additional information the bifunctional dCTP deaminase:dUTPase shows very similar affinities for dCTP and deoxyuridine triphosphate as substrates, overview Mycobacterium tuberculosis H37Rv ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.6.1.23 ? x * 25000, recombinant enzyme, SDS-PAGE Mycobacterium tuberculosis

Synonyms

EC Number Synonyms Comment Organism
3.5.4.13 dCTP deaminase:deoxyuridine triphosphatase
-
 Mycobacterium tuberculosis
3.5.4.13 dCTP deaminase:dUTPase
-
 Mycobacterium tuberculosis
3.5.4.13 deoxycytidine triphosphate deaminase
-
 Mycobacterium tuberculosis
3.6.1.23 dCTP deaminase:dUTPase
-
 Mycobacterium tuberculosis
3.6.1.23 dUTPase
-
 Mycobacterium tuberculosis