Literature summary for 3.2.2.27 extracted from

Knaevelsrud, I.; Ruoff, P.; Anensen, H.; Klungland, A.; Bjelland, S.; Birkeland, N.K.
Excision of uracil from DNA by the hyperthermophilic Afung protein is dependent on the opposite base and stimulated by heat-induced transition to a more open structure (2001), Mutat. Res., 487, 173-190.

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Cloned(Commentary)

Cloned (Comment)	Organism
overexpression in Escherichia coli	Archaeoglobus fulgidus

Inhibitors

Inhibitors	Comment	Organism	Structure
4-hydroxymercuribenzoate	1 mM, 94% inhibition	Archaeoglobus fulgidus
Uracil	5 mM, 51% inhibition	Archaeoglobus fulgidus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.000536	-	uracil-mismatched single-stranded DNA	pH 7.5, 95°C	Archaeoglobus fulgidus
0.00134	-	uracil-mismatched double-stranded DNA	pH 7.5, 70°C	Archaeoglobus fulgidus
0.00151	-	uracil-mismatched double-stranded DNA	pH 7.5, 80°C	Archaeoglobus fulgidus

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
27000	-	gel filtration	Archaeoglobus fulgidus

Organism

Organism	UniProt	Comment	Textmining
Archaeoglobus fulgidus	O28007	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Archaeoglobus fulgidus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.009	-	pH 7.5, 80°C	Archaeoglobus fulgidus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	the protein is devoid of methyl purine and formamidopyrimidine DNA glycosylase activity, respectively	Archaeoglobus fulgidus	?	-	?
uracil-mismatched double-stranded DNA + H2O	the enzyme exhibits opposite base-dependent excision of uracil in order of decreasing efficiency: uridine, U:T, U:C, U:G, U:A	Archaeoglobus fulgidus	uracil + double-stranded DNA with abasic site	-	?
uracil-mismatched single-stranded DNA + H2O	-	Archaeoglobus fulgidus	uracil + single-stranded DNA with abasic site	-	?

Synonyms

Synonyms	Comment	Organism
Afung protein	-	Archaeoglobus fulgidus

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
80	90	-	Archaeoglobus fulgidus

Temperature Range [°C]

Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
20	50	fluorescence and activity measurements following incubation at different temperatures suggest the following model describing structure-activity relationships: at temperatures from 20 to 50°C the protein exists as a compact protein exhibiting low enzyme activity, whereas at temperatures above 50°C, the Afung conformation opens up, which is associated with the acquisition of high enzyme activity	Archaeoglobus fulgidus
65	100	65°C: about 55% of maximal activity, 100°C: about 50% of maximal activity	Archaeoglobus fulgidus

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
70	-	20 min, enzyme in crude protein extract is stable	Archaeoglobus fulgidus

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
additional information	-	additional information	kinetic parameters determined for Afung protein suggest a significantly lower level of enzymatic uracil release in Aeroglobus fulgidus as compared to the mesophilic Escherichia coli	Archaeoglobus fulgidus
0.0175	-	uracil-mismatched double-stranded DNA	pH 7.5, 70°C	Archaeoglobus fulgidus
0.02	-	uracil-mismatched double-stranded DNA	pH 7.5, 80°C	Archaeoglobus fulgidus
0.083	-	uracil-mismatched single-stranded DNA	pH 7.5, 95°C	Archaeoglobus fulgidus

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
4.8	-	-	Archaeoglobus fulgidus
7.5	-	assay at	Archaeoglobus fulgidus

pH Range

pH Minimum	pH Maximum	Comment	Organism
4	7	pH 4.0: about 25% of maximal activity, pH 7.0: about 25% of maximal activity	Archaeoglobus fulgidus

pI Value

Organism	Comment	pI Value Maximum	pI Value
Archaeoglobus fulgidus	isoelectric focusing	-	5.7

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
13.1	-	uracil-mismatched double-stranded DNA	pH 7.5, 70°C	Archaeoglobus fulgidus
13.2	-	uracil-mismatched double-stranded DNA	pH 7.5, 80°C	Archaeoglobus fulgidus
154.9	-	uracil-mismatched single-stranded DNA	pH 7.5, 95°C	Archaeoglobus fulgidus

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Release 2023.1 (January 2023)