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Literature summary for 3.2.2.27 extracted from

  • Engstrom, L.; Partington, O.; David, S.
    An iron-sulfur cluster loop motif in the Archaeoglobus fulgidus uracil-DNA glycosylase mediates efficient uracil recognition and removal (2012), Biochemistry, 51, 5187-5197.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.2.2.27

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Archaeoglobus fulgidus

Protein Variants

Protein Variants Comment Organism
C101A absence of a fully coordinated [4Fe-4S]2+ cluster, loses considerable activity after incubation in assay buffer for two min, with about 20% of the original level of activity remaining Archaeoglobus fulgidus
C85A absence of a fully coordinated [4Fe-4S]2+ cluster, loses considerable activity after incubation in assay buffer for two min, with about 20% of the original level of activity remaining Archaeoglobus fulgidus
K100A absence of a fully coordinated [4Fe-4S]2+ cluster, 3fold and 2fold increase in catalytic efficiency (kcat/Km) for the mutant over wild-type enzyme for double-stranded and single-stranded substrates, respectively Archaeoglobus fulgidus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information significantly different Km-values for the two substrates, with values of 70 nM for double-stranded DNA and 11 nM for single-stranded DNA Archaeoglobus fulgidus
0.000008
-
 uracil-containing single-stranded DNA pH 7.5, 52°C, mutant enzyme K100A Archaeoglobus fulgidus
0.000011
-
 uracil-containing single-stranded DNA pH 7.5, 52°C, wild-type enzyme Archaeoglobus fulgidus
0.000054
-
 uracil-containing double-stranded DNA pH 7.5, 52°C, mutant enzyme K100A Archaeoglobus fulgidus
0.00007
-
 uracil-containing double-stranded DNA pH 7.5, 52°C, wild-type enzyme Archaeoglobus fulgidus

Organism

Organism UniProt Comment Textmining
Archaeoglobus fulgidus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Archaeoglobus fulgidus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
uracil-containing double-stranded DNA + H2O the rate of catalytic turnover (kcat), is higher with double-stranded substrate compared to single-stranded substrate (20/min versus 11/min), respectively. Catalytic efficiency is 3.5fold higher for single-stranded DNA due to the smaller Km-value Archaeoglobus fulgidus uracil + double-stranded DNA with abasic site
-
 ?
uracil-containing single-stranded DNA + H2O the rate of catalytic turnover (kcat), is higher with double-stranded substrate compared to single-stranded substrate (20/min versus 11/min), respectively. Catalytic efficiency is 3.5fold higher for single-stranded DNA due to the smaller Km-value Archaeoglobus fulgidus uracil + single-stranded DNA with abasic site
-
 ?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
additional information
-
 additional information rate constants of uracil excision determined for AFUDG and pseudo-FCL mutants using single-stranded uracil containing or double-stranded U:G base pair-containing substrates under single-turnover conditions Archaeoglobus fulgidus
0.18
-
 uracil-containing single-stranded DNA pH 7.5, 52°C, wild-type enzyme Archaeoglobus fulgidus
0.27
-
 uracil-containing single-stranded DNA pH 7.5, 52°C, mutant enzyme K100A Archaeoglobus fulgidus
0.33
-
 uracil-containing double-stranded DNA pH 7.5, 52°C, wild-type enzyme Archaeoglobus fulgidus
0.85
-
 uracil-containing double-stranded DNA pH 7.5, 52°C, mutant enzyme K100A Archaeoglobus fulgidus

pH Optimum

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

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
additional information
-
 additional information higher catalytic efficiency (kcat/Km) towards single-stranded DNA due to the smaller Km value Archaeoglobus fulgidus
4714
-
 uracil-containing double-stranded DNA pH 7.5, 52°C, wild-type enzyme Archaeoglobus fulgidus
15740
-
 uracil-containing double-stranded DNA pH 7.5, 52°C, mutant enzyme K100A Archaeoglobus fulgidus
16360
-
 uracil-containing single-stranded DNA pH 7.5, 52°C, wild-type enzyme Archaeoglobus fulgidus
33750
-
 uracil-containing single-stranded DNA pH 7.5, 52°C, mutant enzyme K100A Archaeoglobus fulgidus