Information on EC 3.2.2.31 - adenine glycosylase

adenine DNA glycosylase

adenine glycosylase

4 entries

adenine-DNA glycosylase

-

751796

adenine/guanine-specific DNA glycosylase

-

750812

DNA repair adenine glycosylase

-

MicA

-

mutY

MUTYH

-

MYH

hydrolyses free adenine bases from 7,8-dihydro-8-oxoguanine:adenine mismatched double-stranded DNA, leaving an apurinic site.

-

adenine-DNA deoxyribohydrolase (adenine-releasing)

The enzyme serves as a mismatch repair enzyme that works to correct 7,8-dihydro-8-oxoguanine:adenine mispairs that arise in DNA when error-prone synthesis occurs past 7,8-dihydro-8-oxoguanine (GO) lesions in DNA. The enzyme excises the adenine of the mispair, producing an apurinic site sensitive to AP endonuclease activity. After removing the undamaged adenine the enzyme remains bound to the site to prevent EC 3.2.2.23 (MutM) from removing the GO lesion, which could lead to a double strand break. In vitro the enzyme is also active with adenine:guanine, adenine:cytosine, and adenine:7,8-dihydro-8-oxoadenine (AO) mispairs, removing the adenine in all cases.

DNA containing a 5-hydroxyuracil/adenine mismatch + H2O

?

-

Substrates: the enzyme has a moderate affinity for DNA containing a 5-hydroxyuracil/adenine mismatch
Products: -

?

DNA containing a 7,8-dihydro-8-oxo-2'-deoxyguanosine:adenine mismatch + H2O

?

13 entries

DNA containing a 7,8-dihydro-8-oxo-2'-deoxyguanosine:adenine mispair + H2O

?

-

744300, 744299

Substrates: -
Products: -

?

DNA containing a 7,8-dihydro-8-oxo-2'-deoxyguanosine:guanine mismatch + H2O

?

-

745274

Substrates: -
Products: -

?

DNA containing a 7,8-dihydro-8-oxo-2'-deoxyguanosine:thymine mismatch + H2O

?

-

Substrates: low activity
Products: -

?

DNA containing a cytosine:adenine mismatch + H2O

?

-

Substrates: -
Products: -

?

DNA containing a cytosine:adenine mispair + H2O

?

-

744299

Substrates: -
Products: -

?

DNA containing a guanine:adenine mismatch + H2O

?

10 entries

DNA containing a guanine:adenine mispair + H2O

?

-

744300, 744299

Substrates: -
Products: -

?

DNA containing a thymine:guanine mismatch + H2O

?

-

Substrates: low activity
Products: -

?

DNA containing an adenine:7,8-dihydro-8-oxoadenine mismatch + H2O

?

-

Substrates: -
Products: -

?

DNA containing an adenine:7,8-dihydro-8-oxoadenine mismatch + H2O

adenine + DNA with apurinic site

DNA containing an adenine:7,8-dihydro-8-oxoguanine mismatch + H2O

?

DNA containing an adenine:7,8-dihydro-8-oxoguanine mispair + H2O

DNA containing an abasic site + adenine

-

Substrates: primary substrate in vivo
Products: -

?

DNA containing an adenine:8-oxoguanine mismatch + H2O

?

Substrates: -
Products: -

?

DNA containing an adenine:cytosine mismatch + H2O

?

-

Substrates: -
Products: -

?

DNA containing an adenine:guanine mismatch + H2O

?

DNA containing an adenine:guanine mismatch + H2O

adenine + DNA with apurinic site

additional information

?

-

18 entries

DNA containing a 7,8-dihydro-8-oxo-2'-deoxyguanosine:adenine mismatch + H2O

?

11 entries

DNA containing a 7,8-dihydro-8-oxo-2'-deoxyguanosine:adenine mispair + H2O

?

-

744300, 744299

Substrates: -
Products: -

?

DNA containing a cytosine:adenine mismatch + H2O

?

-

Substrates: -
Products: -

?

DNA containing a guanine:adenine mismatch + H2O

?

5 entries

DNA containing an adenine:7,8-dihydro-8-oxoadenine mismatch + H2O

adenine + DNA with apurinic site

DNA containing an adenine:7,8-dihydro-8-oxoguanine mismatch + H2O

?

752171

Substrates: very high specificity
Products: -

?

DNA containing an adenine:7,8-dihydro-8-oxoguanine mispair + H2O

DNA containing an abasic site + adenine

-

Substrates: primary substrate in vivo
Products: -

?

DNA containing an adenine:8-oxoguanine mismatch + H2O

?

Substrates: -
Products: -

?

DNA containing an adenine:guanine mismatch + H2O

adenine + DNA with apurinic site

additional information

?

-

[4Fe-4S]-center

Zinc

MUTYH has a Zn-binding motif in a interdomain connector region. The Zn-binding motif comprises residues His56, Cys300, Cys307 and Cys310. The interdomain connector region, including the Zn-binding motif, is exposed on the MUTYH surface

774803

heparin

-

competitive inhibitor

750812

suramin

-

competitive inhibitor

750812

0.000224

DNA containing a 5-hydroxyuracil/adenine mismatch

-

at pH 7.6 and 37°C

-

0.000025

DNA containing a 5-hydroxyuracil/adenine mismatch

-

at pH 7.6 and 37°C

-

1.07

DNA containing a 5-hydroxyuracil/adenine mismatch

-

at pH 7.6 and 37°C

-

-

744395, 745065, 750812

-

Corynebacterium pseudotuberculosis ADK29561.1

-

-

-

-

8 entries

-

745928, 751044, 773676

UniProt

-

-

-

-

Schizosaccharomyces pombe

-

-

-

SwissProt

-

UniProt

-

745780

-

Highest Expressing Human Cell Lines

Filter by:

Cell Line Links

Gene Links

malfunction

Schizosaccharomyces pombe

-

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

745780

metabolism

physiological function

9 entries

additional information

Mus musculus

Q99P21

8-oxoguanine is recognized by residues Gln110, Thr111, Leu148 and Tyr150. MUTYH recognizes N6-H of adenine by a hydrogen bond with Oepsilon of the polar side chain of Gln251

?

homodimer

-

mutant enzyme C199H, hanging drop vapor diffusion method, using 1.6-1.8 M ammonium sulfate and 100 mM Tris-HCl, pH 8.5

744297

enzyme bound to an azaribose transition state analog

745928

in complex with DNA, hanging drop vapor diffusion method, using 100 mM Tris, pH 8.0, 250 mM NaOAc, 29% (w/v) PEG 4000

crystal structures of complexes between mouse MUTYH and DNA, and between the C-terminal domain of mouse MUTYH and human proliferating cell nuclear antigen PCNA

Mus musculus

Q99P21

774803

D179A

-

active site mutant

E78A

-

active site mutant

E78A/D179A

-

active site mutant

C199H

the mutant shows wild type activity

744297

C199S

the mutant shows wild type activity

D138C

D138E

the mutant shows wild type activity

D138N

E37C

E37D

the mutant shows 68% of wild type activity

E37Q

inactive

E37S

inactive

G253A

-

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

-

the mutant has a reduced adenine glycosylase activity and affinity for substrate analogues compared to the wild type enzyme

K142A

4 entries

K157A

-

the mutation has no effect on enzyme activities

K158A

-

the mutation has no effect on enzyme activities

K196A

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

744153

K198A

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

744153

K20A

-

the mutation still catalyzes beta- and delta-elimination reactions and can be trapped as covalent enzyme-DNA intermediates by chemical reduction

745274

Q182L

-

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

the mutant shows wild type activity

744153

S120K

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

744295

S120K/K142A

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

744295

V45A

-

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

-

the mutant has similar binding affinities to thymine/guanine as the wild type enzyme

Y82C

-

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

-

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

-

the mutant is unable to discriminate between guanine and 7,8-dihydro-8-oxo-2'-deoxyguanosine when paired with 2'-deoxy-2'-fluoroadenosine

D144N

active site mutant

P164C

active site mutant

Y126F

the mutant shows a 260fold slower rate of adenine removal compared to the wild type enzyme

745928

G382D

-

the mutant enzyme shows a reduced ability to recognize and repair 7,8-dihydro-8-oxo-2'-deoxyguanosine:adenine mismatches

Y165C

-

the mutant enzyme shows a reduced ability to recognize and repair 7,8-dihydro-8-oxo-2'-deoxyguanosine:adenine mismatches

D149A

-

active site mutant

E48A

-

active site mutant

E48A/D149A

-

active site mutant

Schizosaccharomyces pombe

F415A/S416A

Mus musculus

Q99P21

mutantions strongly reduce the glycosylase activity for A:8-oxoG mispairing

774803

D172N

-

inactive

745780

45

-

melting temperature

-

ammonium sulfate precipitation and HiTrap heparin column chromatography

-

ammonium sulfate precipitation, Ni-NTA column chromatography, and Superdex 75 gel filtration

-

ammonium sulfate precipitation, phosphocellulose column chromatography, hydroxylapatite column chromatography, heparin column chromatography, and Hitrap-S column chromatography

-

heparin-Sepharose column chromatography

-

745913

HiTrap heparin column chromatography

-

MonoQ column chromatography

Ni-NTA column chromatography

Q-Sepharose and SP-Sepharose column chromatography, and Superdex 200 gel filtration

-

745259

Q-Sepharose column chromatography and HiTrap chelating column chromatography

-

-

expressed as chimeric glycosylase in Escherichia coli JM101 cells

expressed in Escherichia coli BL21 (DE3) pLysS cells

-

expressed in Escherichia coli BL21 Rosetta pLys and PR70 cells

Helicobacter pylori

-

744766

expressed in Escherichia coli BL21(DE3) cells

expressed in Escherichia coli CC104 cells

-

745274

expressed in Escherichia coli JM101 cells

-

648678, 745913

expressed in Escherichia coli JM101, CC104 and GT100 cells

-

expressed in Escherichia coli PR70(DE3) cells

-

expressed in Escherichia coli PR70/DE3 cells

-

expressed in HEK-293 cells

Rattus norvegicus

Q8R5G2

745914

expressed in mutY gene deficient Escherichia coli BH980 cells