Literature summary for 3.2.2.27 extracted from

Moe, E.; Leiros, I.; Smalas, A.O.; McSweeney, S.
The crystal structure of mismatch-specific uracil-DNA glycosylase (MUG) from Deinococcus radiodurans reveals a novel catalytic residue and broad substrate specificity (2006), J. Biol. Chem., 281, 569-577.

Cloned(Commentary)

Cloned (Comment)	Organism
MUG, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression of wild-type and mutant enzymes in Escherichia coli strain Bl21(DE3)	Deinococcus radiodurans

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant wild-type MUG and MUG mutant D93A, crystal growth by mixing 0.001 ml drops of 10 mg/ml protein with a solution containing 0.2 M sodium acetate trihydrate, 0.1 M sodium cacodylate, pH 6.5, and 30% w/v polyethylene glycol 8000, equilibration at 18°C, hexagonal crystals suitable for data collection purposes appear after 1 day, X-ray diffraction structure determination and analysis at 1.7-1.75 A resolution, molecular replacement	Deinococcus radiodurans

Crystallization (Comment)

Organism

purified recombinant wild-type MUG and MUG mutant D93A, crystal growth by mixing 0.001 ml drops of 10 mg/ml protein with a solution containing 0.2 M sodium acetate trihydrate, 0.1 M sodium cacodylate, pH 6.5, and 30% w/v polyethylene glycol 8000, equilibration at 18°C, hexagonal crystals suitable for data collection purposes appear after 1 day, X-ray diffraction structure determination and analysis at 1.7-1.75 A resolution, molecular replacement

Deinococcus radiodurans

Protein Variants

Protein Variants	Comment	Organism
D93A	site-directed mutagenesis, inactive mutant	Deinococcus radiodurans

Organism

Organism	UniProt	Comment	Textmining
Deinococcus radiodurans	Q9RWH9	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3)	Deinococcus radiodurans

Reaction

Reaction	Comment	Organism	Reaction ID
Hydrolyses single-stranded DNA or mismatched double-stranded DNA and polynucleotides, releasing free uracil	catalytic reaction mechanism, the catalytic residue is Asp93	Deinococcus radiodurans	a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.01	-	purified recombinant MUG	Deinococcus radiodurans

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
uracil-mismatched double-stranded DNA + H2O	30-bp dsDNA oligonucleotide substrates containing G-C, G-U, G-T, and A-U base pairs. Determination of kinetics using commercially available nick-translated calf thymus DNA with deoxy[5-3H]uridine 5'-triphosphate as substrate. To perform efficient glycoside bond cleavage, drMUG must stabilize the mismatched uracil in the specificity pocket, nucleotide stabilization by Tyr46, substrate binding mechanism, overview. Binding of thymine in the activity pocket is probably prevented by Ser36 and Ser39 in MUG, binding of cytosine is prevented by Asp84	Deinococcus radiodurans	uracil + double-stranded DNA with abasic site	-	?
uracil-mismatched single-stranded DNA + H2O	single-stranded DNA containing uracil labeled with fluorescein in the 5'-end, sequence overview. Determination of kinetics using commercially available nick-translated calf thymus DNA with deoxy[5-3H]uridine 5'-triphosphate as substrate. To perform efficient glycoside bond cleavage, drMUG must stabilize the mismatched uracil in the specificity pocket, nucleotide stabilization by Tyr46, substrate binding mechanism, overview. Binding of thymine in the activity pocket is probably prevented by Ser36 and Ser39 in MUG, binding of cytosine is prevented by Asp84	Deinococcus radiodurans	uracil + single-stranded DNA with abasic site	-	?

Synonyms

Synonyms	Comment	Organism
mismatch-specific uracil-DNA glycosylase	-	Deinococcus radiodurans
More	three MUGs form a distintc MUG/thymidine-DNA glycosylase subfamily, called the MUG2 family	Deinococcus radiodurans
MUG	-	Deinococcus radiodurans

Temperature Optimum [°C]

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

pH Optimum

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