Literature summary for 3.6.1.23 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.

Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment)	Organism
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

Crystallization (Comment)

Organism

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

Inhibitors	Comment	Organism	Structure
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]

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

Molecular Weight [Da]

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

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
dUTP + H2O	Mycobacterium tuberculosis	-	dUMP + diphosphate	-	?
dUTP + H2O	Mycobacterium tuberculosis H37Rv	-	dUMP + diphosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Mycobacterium tuberculosis	P9WNS5	-	-
Mycobacterium tuberculosis H37Rv	P9WNS5	-	-

Purification (Commentary)

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

Reaction

Reaction	Comment	Organism	Reaction ID
dUTP + H2O = dUMP + diphosphate	catalytic mechanism	Mycobacterium tuberculosis	a

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

Synonyms	Comment	Organism
dCTP deaminase:dUTPase	-	Mycobacterium tuberculosis
dUTPase	-	Mycobacterium tuberculosis