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Literature summary for 3.2.2.27 extracted from

  • Raeder, I.L.; Moe, E.; Willassen, N.P.; Smalas, A.O.; Leiros, I.
    Structure of uracil-DNA N-glycosylase (UNG) from Vibrio cholerae: mapping temperature adaptation through structural and mutational analysis (2010), Acta Crystallogr. Sect. F, 66, 130-136.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
vcUNG, expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) Vibrio cholerae

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant His-tagged enzyme, hanging drop vapour diffusion, mixing of 0.001 ml of protein solution containing 15 mg/ml protein in 50 mM Tris-HCl, pH 7.5, and 5 mM 2-mercaptoethanol, with 0.001 ml of reservoir solution containing 50 mM Na MES, pH 6.0, and 28% w/v PEG 3350, equilibration against a reservoir containing 1.0 ml precipitant solution, room temperature, 1 week, X-ray diffraction structure determination and analysis at 1.5 A resolution, modelling Vibrio cholerae

Protein Variants

Protein Variants Comment Organism
H194R site-directed mutagenesis, the mutant shows altered kinetics compared to the wild-type enzyme Vibrio cholerae
V90R site-directed mutagenesis, the mutant shows altered kinetics compared to the wild-type enzyme Vibrio cholerae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0004
-
dUMP DNA pH 7.5, recombinant wild-type enzyme Aliivibrio salmonicida
0.0011
-
dUMP DNA pH 7.5, recombinant mutant V90R Vibrio cholerae
0.0018
-
dUMP DNA pH 7.5, recombinant mutant H194R Vibrio cholerae
0.0028
-
dUMP DNA pH 7.5, recombinant wild-type enzyme Vibrio cholerae

Organism

Organism UniProt Comment Textmining
Aliivibrio salmonicida
-
-
-
Vibrio cholerae Q9KPK8
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration Vibrio cholerae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
dUMP DNA + H2O
-
Vibrio cholerae dUMP + DNA
-
?
dUMP DNA + H2O
-
Aliivibrio salmonicida dUMP + DNA
-
?

Subunits

Subunits Comment Organism
More asUNG structure homology modelling based on the crystal structure of vcUNG from Vibrio cholerae, asUNG possesses typical cold-adapted features compared with vcUNG, overview Aliivibrio salmonicida
More UNG structure modelling, overview Vibrio cholerae

Synonyms

Synonyms Comment Organism
More the enzyme belongs to the uracil DNA glycosylase family Vibrio cholerae
More the enzyme belongs to the uracil DNA glycosylase family Aliivibrio salmonicida
UNG
-
Vibrio cholerae
UNG
-
Aliivibrio salmonicida
uracil-DNA N-glycosylase
-
Vibrio cholerae
uracil-DNA N-glycosylase
-
Aliivibrio salmonicida

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
154
-
dUMP DNA pH 7.5, recombinant mutant V90R Vibrio cholerae
174
-
dUMP DNA pH 7.5, recombinant wild-type enzyme Aliivibrio salmonicida
293
-
dUMP DNA pH 7.5, recombinant wild-type enzyme Vibrio cholerae
821
-
dUMP DNA pH 7.5, recombinant mutant H194R Vibrio cholerae

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Vibrio cholerae
7.5
-
assay at Aliivibrio salmonicida