Information on EC 3.2.2.31 - adenine glycosylase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
3.2.2.31
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RECOMMENDED NAME
GeneOntology No.
adenine glycosylase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
hydrolyses free adenine bases from 7,8-dihydro-8-oxoguanine:adenine mismatched double-stranded DNA, leaving an apurinic site.
show the reaction diagram
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-
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SYSTEMATIC NAME
IUBMB Comments
adenine-DNA deoxyribohydrolase (adenine-releasing)
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
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an enzyme-knockout strain displays a 36fold higher frequency of spontaneous mutations than the wild type strain. Enzyme gene disruption causes increased sensitivity to H2O2 but not UV irradiation
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
DNA containing a 5-hydroxyuracil/adenine mismatch + H2O
?
show the reaction diagram
-
the enzyme has a moderate affinity for DNA containing a 5-hydroxyuracil/adenine mismatch
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-
?
DNA containing a 7,8-dihydro-8-oxo-2'-deoxyguanosine:adenine mismatch + H2O
?
show the reaction diagram
DNA containing a 7,8-dihydro-8-oxo-2'-deoxyguanosine:adenine mispair + H2O
?
show the reaction diagram
-
-
-
-
?
DNA containing a 7,8-dihydro-8-oxo-2'-deoxyguanosine:guanine mismatch + H2O
?
show the reaction diagram
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-
-
-
?
DNA containing a 7,8-dihydro-8-oxo-2'-deoxyguanosine:thymine mismatch + H2O
?
show the reaction diagram
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low activity
-
-
?
DNA containing a cytosine:adenine mismatch + H2O
?
show the reaction diagram
-
-
-
-
?
DNA containing a cytosine:adenine mispair + H2O
?
show the reaction diagram
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-
-
-
?
DNA containing a guanine:adenine mismatch + H2O
?
show the reaction diagram
DNA containing a guanine:adenine mispair + H2O
?
show the reaction diagram
-
-
-
-
?
DNA containing a thymine:guanine mismatch + H2O
?
show the reaction diagram
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low activity
-
-
?
DNA containing an adenine:7,8-dihydro-8-oxoadenine mismatch + H2O
?
show the reaction diagram
-
-
-
-
?
DNA containing an adenine:7,8-dihydro-8-oxoguanine mispair + H2O
DNA containing an abasic site + adenine
show the reaction diagram
-
primary substrate in vivo
-
-
?
DNA containing an adenine:cytosine mismatch + H2O
?
show the reaction diagram
-
-
-
-
?
DNA containing an adenine:guanine mismatch + H2O
?
show the reaction diagram
-
-
-
-
?
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
DNA containing a 7,8-dihydro-8-oxo-2'-deoxyguanosine:adenine mismatch + H2O
?
show the reaction diagram
DNA containing a 7,8-dihydro-8-oxo-2'-deoxyguanosine:adenine mispair + H2O
?
show the reaction diagram
-
-
-
-
?
DNA containing a cytosine:adenine mismatch + H2O
?
show the reaction diagram
-
-
-
-
?
DNA containing a guanine:adenine mismatch + H2O
?
show the reaction diagram
DNA containing an adenine:7,8-dihydro-8-oxoguanine mispair + H2O
DNA containing an abasic site + adenine
show the reaction diagram
-
primary substrate in vivo
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?
additional information
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
[4Fe-4S]-center
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000224
DNA containing a 5-hydroxyuracil/adenine mismatch
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at pH 7.6 and 37°C
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000025
DNA containing a 5-hydroxyuracil/adenine mismatch
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at pH 7.6 and 37°C
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.07
DNA containing a 5-hydroxyuracil/adenine mismatch
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at pH 7.6 and 37°C
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SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
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Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
mutant enzyme C199H, hanging drop vapor diffusion method, using 1.6-1.8 M ammonium sulfate and 100 mM Tris-HCl, pH 8.5
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enzyme bound to an azaribose transition state analog
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45
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melting temperature
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation and HiTrap heparin column chromatography
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ammonium sulfate precipitation, Ni-NTA column chromatography, and Superdex 75 gel filtration
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ammonium sulfate precipitation, phosphocellulose column chromatography, hydroxylapatite column chromatography, heparin column chromatography, and Hitrap-S column chromatography
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heparin-Sepharose column chromatography
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HiTrap heparin column chromatography
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Ni-NTA column chromatography
Q-Sepharose and SP-Sepharose column chromatography, and Superdex 200 gel filtration
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21 (DE3) pLysS cells
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expressed in Escherichia coli BL21 Rosetta pLys and PR70 cells
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expressed in Escherichia coli BL21(DE3) cells
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expressed in Escherichia coli CC104 cells
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expressed in Escherichia coli JM101 cells
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expressed in Escherichia coli JM101, CC104 and GT100 cells
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expressed in Escherichia coli PR70(DE3) cells
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expressed in Escherichia coli PR70/DE3 cells
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expressed in HEK-293 cells
expressed in mutY gene deficient Escherichia coli BH980 cells
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C199H
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the mutant shows wild type activity
C199S
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the mutant shows wild type activity
D138E
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the mutant shows wild type activity
D138N
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inactive
E37D
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the mutant shows 68% of wild type activity
E37Q
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inactive
E37S
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inactive
G253A
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when 2'-deoxy-2'-fluoroadenosine in duplex paired with guanine or 7,8-dihydro-8-oxo-2'-deoxyguanosine is used, the mutant shows reduced binding affinity. Additionally, compromised glycosylase activity of the mutant enzyme is observed using the nonoptimal guanine:adenine substrate, or at low reaction temperatures
G253D
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the mutant has a reduced adenine glycosylase activity and affinity for substrate analogues compared to the wild type enzyme
K157A
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the mutation has no effect on enzyme activities
K158A
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the mutation has no effect on enzyme activities
K196A
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the mutant enzyme exhibits a slight reduction of the rate of adenine removal from a G:A base pair-containing duplex compared to the wild type enzyme
K198A
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the mutant enzyme exhibits a significant reduction (15fold) of the rate of adenine removal from a G:A base pair-containing duplex compared to the wild type enzyme
K20A
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the mutation still catalyzes beta- and delta-elimination reactions and can be trapped as covalent enzyme-DNA intermediates by chemical reduction
Q182L
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the mutant has significantly lower binding and glycosylase activities for adenine:guanine and adenine:7,8-dihydro-8-oxo-guanine mismatches than the wild type enzyme. The mutant has substantially increased affinity towards thymine:guanine, however, it does not exhibit any thymine/guanine or thymine/7,8-dihydro-8-oxo-guanine glycosylase activity
R194A
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the mutant shows wild type activity
S120K
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the mutant with bifunctional glycosylase/AP lyase activity is capable of catalyzing DNA strand scission at a rate equivalent to that of adenine excision for both guanine:adenine and 7,8-dihydro-8-oxo-2'-deoxyguanosine:adenine mispair substrates
S120K/K142A
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the mutant with bifunctional glycosylase/AP lyase activity is capable of catalyzing DNA strand scission at a rate equivalent to that of adenine excision for both guanine:adenine and 7,8-dihydro-8-oxo-2'-deoxyguanosine:adenine mispair substrates
V45A
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the mutant has significantly lower binding and glycosylase activities for adenine:guanine and adenine:7,8-dihydro-8-oxo-guanine mismatches than the wild type enzyme. The mutant has substantially increased affinity towards thymine:guanine, however, it does not exhibit any thymine/guanine or thymine/7,8-dihydro-8-oxo-guanine glycosylase activity
V45A/Q182L
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the mutant has similar binding affinities to thymine/guanine as the wild type enzyme
Y82C
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the mutant has a reduced adenine glycosylase activity and affinity for substrate analogues compared to the wild type enzyme. Additionally, compromised glycosylase activity of the mutant enzyme is observed using the nonoptimal guanine:adenine substrate, or at low reaction temperatures
Y82F
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when 2’-deoxy-2’-fluoroadenosine in duplex paired with guanine or 7,8-dihydro-8-oxo-2'-deoxyguanosine is used, the mutant shows reduced binding affinity. Additionally, compromised glycosylase activity of the mutant enzyme is observed using the nonoptimal guanine:adenine substrate, or at low reaction temperatures
Y82L
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the mutant is unable to discriminate between guanine and 7,8-dihydro-8-oxo-2'-deoxyguanosine when paired with 2'-deoxy-2'-fluoroadenosine
Y126F
the mutant shows a 260fold slower rate of adenine removal compared to the wild type enzyme
G382D
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the mutant enzyme shows a reduced ability to recognize and repair 7,8-dihydro-8-oxo-2'-deoxyguanosine:adenine mismatches
Y165C
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the mutant enzyme shows a reduced ability to recognize and repair 7,8-dihydro-8-oxo-2'-deoxyguanosine:adenine mismatches