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Literature summary for 3.2.2.27 extracted from

  • Leiros, I.; Moe, E.; Smala, A.O.; McSweeney, S.
    Structure of the uracil-DNA N-glycosylase (UNG) from Deinococcus radiodurans (2005), Acta Crystallogr. Sect. D, D61, 1049-1056.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene dr0689, expression of His-tagged UNG type 1 in Escherichia coli strain BL21(DE3) Deinococcus radiodurans

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant His-tagged UNG type 1, hanging drop vapour diffusion method, 0.001 ml of 10 mg/ml protein in 70 mM Tris-HCl, 10 mM NaCl, 1 mM EDTA pH 8.0, 100 mg/ml bovine serum albumin, are mixed with 0.001 ml of precipitant solution containing 0.2 M ammonium nitrate and 17% w/v PEG 3000 at 4°C for a few days, 20% v/v glycerol as cryoprotectant, X-ray diffraction structure determination and analysis at 1.8-1.9 A resolution Deinococcus radiodurans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetics Deinococcus radiodurans
0.0007
-
uracil-mismatched double-stranded DNA pH 8.0, 37°C, recombinant enzyme Deinococcus radiodurans

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
27700
-
x * 27700, about, sequence calculation Deinococcus radiodurans

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
uracil-mismatched double-stranded DNA + H2O Deinococcus radiodurans the enzyme removes uracil from DNA, which can occur by misincorporation of dUMP in place of dTMP during DNA synthesis or by deamination of cytosine, resulting in U-A or U-G mispairs uracil + double-stranded DNA with abasic site
-
?

Organism

Organism UniProt Comment Textmining
Deinococcus radiodurans Q9RWH9 gene dr0689 encoding the type 1 UDG, i.e. UNG
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged UNG type 1 from Escherichia coli strain BL21(DE3) by affinity chromatography Deinococcus radiodurans

Reaction

Reaction Comment Organism Reaction ID
Hydrolyses single-stranded DNA or mismatched double-stranded DNA and polynucleotides, releasing free uracil uracil-DNA glycosylases are widespread enzymes that are found in all living organisms. UDGs form a central part of the DNA-repair machinery since they initiate the DNA base-excision repair pathway by hydrolysing the N-glycosidic bond between uracil and the deoxyribose sugar thereby catalysing the removal of mis-incorporated uracil from DNA Deinococcus radiodurans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
uracil-mismatched double-stranded DNA + H2O the enzyme removes uracil from DNA, which can occur by misincorporation of dUMP in place of dTMP during DNA synthesis or by deamination of cytosine, resulting in U-A or U-G mispairs Deinococcus radiodurans uracil + double-stranded DNA with abasic site
-
?
uracil-mismatched double-stranded DNA + H2O drUNG is able to excise uracil from both U-A and U-G double-stranded DNA and single-stranded DNA, substrate is commercially available nick-translated calf thymus DNA with deoxy[5-3H]uridine 5'-triphosphate: DNA binding, the active-site environment, and uracil specificity, structure-activity relationship, overview Deinococcus radiodurans uracil + double-stranded DNA with abasic site
-
?

Subunits

Subunits Comment Organism
? x * 27700, about, sequence calculation Deinococcus radiodurans
More type 1 UNG structure analysis and comparison with the human and Escherichia coli enzymes, structure-activity relationship, overview Deinococcus radiodurans

Synonyms

Synonyms Comment Organism
dr0689
-
Deinococcus radiodurans
type 1 UDG
-
Deinococcus radiodurans
UDG
-
Deinococcus radiodurans
UNG
-
Deinococcus radiodurans
uracil-DNA N-glycosylase
-
Deinococcus radiodurans

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Deinococcus radiodurans

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
15.58
-
uracil-mismatched double-stranded DNA pH 8.0, 37°C, recombinant enzyme Deinococcus radiodurans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
assay at Deinococcus radiodurans

pI Value

Organism Comment pI Value Maximum pI Value
Deinococcus radiodurans sequence calculation
-
6.97