Information on EC 3.2.2.21 - DNA-3-methyladenine glycosylase II

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

EC NUMBER
COMMENTARY
3.2.2.21
-
RECOMMENDED NAME
GeneOntology No.
DNA-3-methyladenine glycosylase II
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
hydrolysis of alkylated DNA, releasing 3-methyladenine, 3-methylguanine, 7-methylguanine and 7-methyladenine
show the reaction diagram
involved in the removal of alkylated bases from DNA in Escherichia coli (cf. EC 2.1.1.63 methylated-DNA-[protein]-cysteine S-methyltransferase)
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of N-glycosyl bond
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
alkylated-DNA glycohydrolase (releasing methyladenine and methylguanine)
Involved in the removal of alkylated bases from DNA in Escherichia coli (cf. EC 2.1.1.63 methylated-DNA---[protein]-cysteine S-methyltransferase).
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3-methyladenine DNA glycosylase II
-
-
3-methyladenine-DNA glycosidase II
-
-
-
-
3-methyladenine-DNA glycosylase II
-
-
-
-
ALK
Escherichia coli JM105
-
-
-
AlkA
-
-
-
-
AlkA
P04395
-
AlkA protein
Escherichia coli AB1157, Escherichia coli JM105, Escherichia coli MV1161
-
-
-
alkylpurine-DNA-N-glycosylase
-
-
deoxyribonucleate 3-methyladenine glycosidase II
-
-
-
-
DNA glycosylase
-
-
DNA-3-methyladenine glycosidase II
-
-
-
-
helix-hairpin-helix DNA glycosylase
-
-
m3A DNA glycosylase II
-
-
N-methylpurine-DNA glycosylase
-
-
TagII
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
89287-38-7
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
strain ATCC 14579
-
-
Manually annotated by BRENDA team
K12 derivatives
-
-
Manually annotated by BRENDA team
strain BW 9062
-
-
Manually annotated by BRENDA team
tagA mutant BK2114
-
-
Manually annotated by BRENDA team
Escherichia coli AB1157
AB1157
-
-
Manually annotated by BRENDA team
Escherichia coli BW 9062
strain BW 9062
-
-
Manually annotated by BRENDA team
Escherichia coli JM105
JM105
-
-
Manually annotated by BRENDA team
Escherichia coli MV1161
MV1161
-
-
Manually annotated by BRENDA team
human; MDA-MB231
-
-
Manually annotated by BRENDA team
mouse; transgenic mouse lines SVT125, 127 and 248, expressing the SV40 large T-antigen gene
-
-
Manually annotated by BRENDA team
recombinant enzyme
-
-
Manually annotated by BRENDA team
no activity in Escherichia coli
alk mutants
-
-
Manually annotated by BRENDA team
fission yeast
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
P04395
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
calf thymus DNA
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
cellular repair of alkylated DNA base modifications
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
DNA repair enzyme, inducible base excision repair pathway of DNA alkylation damage
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
P04395
DNA repair enzyme, inducible base excision repair pathway of DNA alkylation damage
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
induced by cell exposure to sublethal doses of alkylating agents, important role in the excision of base damage from single-stranded regions transiently formed in DNA during transcription and replication
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
short-patch base excision repair pathway, repair of alkylation and oxidative DNA damage
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
binds mismatch base-pairs in DNA
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
inducible pathway for repair of DNA damaged by simple alkylating agents such as methylmethanesulfonate and N-methyl-N'-nitro-N-nitrosoguanidine
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
Escherichia coli MV1161
-
-, inducible pathway for repair of DNA damaged by simple alkylating agents such as methylmethanesulfonate and N-methyl-N'-nitro-N-nitrosoguanidine
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
Escherichia coli MV1161
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
Escherichia coli JM105
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
Escherichia coli BW 9062
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
Escherichia coli AB1157
-
-, DNA repair enzyme, inducible base excision repair pathway of DNA alkylation damage
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + 7-methyladenine + DNA
show the reaction diagram
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 3-methylguanine + 7-methyladenine + 7-methylguanine + DNA
show the reaction diagram
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 3-methylguanine + 7-methyladenine + 7-methylguanine + DNA
show the reaction diagram
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 3-methylguanine + 7-methylguanine + 7-methyladenine + ?
show the reaction diagram
-
AlkA has significant glycosylase activity towards each of the normal bases in DNA. AlkA binds nonspecifically to DNA and neither mismatches nor 7-methylguanine lesions are specifically recognized in the ground state
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + 3-methylguanine
show the reaction diagram
-
AlkD has high activity towards 7-methylguanine but removes 3-methylguanine more slowly as compared with Escherichia coli AlkA
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + ?
show the reaction diagram
-
-
ratio of 3-methyladenine/7-methylguanine is 29:1 for wild-type enzyme, 24:1 for mutant enzyme N169S and 26:1 for mutant enzyme N169A, no production of 7-methylguanine is detected with mutant enzyme N169D
-
?
alkylated DNA + H2O
?
show the reaction diagram
-
the efficiency of 3-methyladenine and 3-methylguanine removal was 510 times slower for Mag1 than for Escherichia coli AlkA whereas the rate of 7-methylguanine removal is similar to the two enzymes
-
-
?
alkylated DNA + H2O
3-methyladenine + N3-methylcytosine + 1,N6-ethenoadenine + DNA
show the reaction diagram
-
-
excision of N3-methylcytosine is about twice as fast as excision of 3-methyladenine, and the rate for 1,N6-ethenoadenine is further 5-10 times higher
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + DNA
show the reaction diagram
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + hypoxanthine + DNA
show the reaction diagram
-
-
-
-
?
alkylated DNA + H2O
3-methyl-2'-deoxyadenosine + 3-deaza-3-methyl-2'-deoxyadenosine + DNA
show the reaction diagram
-
the sequences of the oligos used in the assay are 5'-CGATAGCATCCTYCCTTCTCTCCAT-3', where Y is the location of the lesion base, and 5'-ATGGAGAGAAGGZAGGATGCTATCG-3' for the complementary strand, where Z is the base opposite the lesion
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + 1,N6-ethenoadenine + hypoxanthine + DNA
show the reaction diagram
-
-
-
-
?
DNA treated with beta-[3H]propiolactone + H2O
N1-(carboxyethyl)adenine + N7-(carboxymethyl)guanine
show the reaction diagram
-
-
-
-
?
duplex oligonucleotide substrate containing ethenoadenine + H2O
ethenoadenine + oligonucleotide
show the reaction diagram
P29372
-
-
-
?
duplex oligonucleotide substrate containing ethenoadenine and hypoxanthine + H2O
ethenoadenine + hypoxanthine + oligonucleotide
show the reaction diagram
-
-
-
-
?
synthetic oligosaccharide + H2O
?
show the reaction diagram
-
synthetic oligosaccharide containing a single ethano or etheno adduct (3,N4-ethanocytosine, 1,N6-ethanoadenine, 3,N4-ethenocytosine or 1,N6-ethenoadenine), 20fold lower excision activity towards 3,N4-ethanocytosine and 1,N6-ethanoadenine than that towards their structurally analogous 3,N4-ethenocytosine or 1,N6-ethenoadenine. The enzyme is capable of excising the ethano base paired with any of the four natural bases
-
-
?
duplex oligonucleotide substrate containing ethenoadenine and hypoxanthine + H2O
ethenoadenine + hypoxanthine + oligonucleotide
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
-
-
-
-
additional information
?
-
-
no activity on O2-methylcytosine, O2-methylthymine, O4-methylthymine or O6-methylguanine
-
-
-
additional information
?
-
-
releases 3-methyladenine, 3-methylguanine, 7-methylguanine, 7-(2-chloroethyl)guanine, 7-(2-hydroxyethyl)guanine, 1,N6-ethenoadenine, hypoxanthine, and guanine from damaged and normal DNA, adenine, thymine, cytosine,3-ethenoguanine, 8-oxoguanine, 7-(2-ethoxyethyl)guanine, O2-methylthymine, and O2-methylcytosine are not released
-
-
-
additional information
?
-
-
removes alkylation damage from duplex and single stranded DNA
-
-
-
additional information
?
-
-
releases 3-methyladenine, 3-methylguanine, 7-methylguanine, O2-methylthymine, O2-methylcytosine, 7-(2-chloroethyl)guanine, 7-(2-hydroxyethyl)guanine, 7-(2-ethoxyethyl)guanine, 1,N6-ethenoadenine, hypoxanthine, adenine, guanine, thymine and cytosine from damaged and normal DNA, 3-ethenoguanine and 8-oxoguanine are not released
-
-
-
additional information
?
-
-
excises N7 guanine adducts completely, also capable of removing normal base residues from DNA
-
-
-
additional information
?
-
-
also capable to relase xanthine and oxanine, guanine lesions induced by nitrosation
-
-
-
additional information
?
-
-
removes N-alkylpurine damage, unrepaired DNA damage leads to carcinogenesis, cell death, and aging, also releases 1,N6-ethenoadenine
-
-
-
additional information
?
-
-
with sulfur mustard treated DNA as substrate enzyme releases 7-hydroxyethylthioethyl guanine and 3-hydroxyethylthioethyl adenine, can also release carboxyethylate purines from DNA
-
-
-
additional information
?
-
-
removes also N-alkylpurines and cyclic ethenoadducta of adenine, guanine and cytosine
-
-
-
additional information
?
-
-
fails to excise N7 guanine adducts
-
-
-
additional information
?
-
-
enzyme cleaves pBR322 and pAlk10 plasmids, excises adducts formed by chloroacetaldehyde, acrolein and croton aldehyde but is not able to excise malionaldehyde and bulky p-benzochinone adducts
-
-
-
additional information
?
-
-
removes also hypoxanthine and 1,N6-ethenoadenine
-
-
-
additional information
?
-
-
enzyme releases N1-(carboxyethyl)adenine and N7-(carboxymethyl)guanine from DNA treated with beta-propiolactone, but does not release the aflatoxin B-1 adduct at N-7 of guanine, release of 3-methyladenine from single-stranded DNA is 9% of the rate from double-stranded DNA, enzyme will not release 2,6-diamino-4-hydroxy-5-(N-methylformamido)pyrimidine, the alkali-induced derivative of 7-methylguanine, in which the imidazole ring is opened
-
-
-
additional information
?
-
-
enzyme also excises thymine residues oxidized in the methyl group like 5-formyluracil and 5-hydroxymethyluracil
-
-
-
additional information
?
-
-
excises alkylated bases only
-
-
-
additional information
?
-
-
repairs 5-formyluracil, a major thymine lesion produced by reactive oxygen radicals and photosensitized oxidation, 5-hydroxymethyluracil, another major thymine methyl oxidation product, is not a substrate
-
-
-
additional information
?
-
-
AlkD is involved exclusively in the repair of alkylation damage
-
-
-
additional information
?
-
-
enzyme is involved in DNA excision repair
-
-
-
additional information
?
-
-
the enzyme is induced in response to DNA alkylation, and it protects cells from alkylated nucleobases by catalyzing their excision
-
-
-
additional information
?
-
-
no detectable affinity for hypoxanthine, 1,N6-ethenoadenine, 8-oxoguanine and 5-formyluracil
-
-
-
additional information
?
-
-
no significant activity is found towards deamination products, ethenoadducts or oxidation products
-
-
-
additional information
?
-
-
AlkD does not excise 1,N6-ethenoadenine
-
-
-
additional information
?
-
-
the enzyme binds preferentially DNA ends, more tightly than hypoxanthine lesions, exhibits significant product inhibition under multiple-turnover conditions, and binds approximately 10fold more tightly to an abasic site than to a hypoxanthine lesion site
-
-
-
additional information
?
-
-
the enzyme has little activity on correctly base-paired adenine. N3-deazaadenine is not substantially cleaved opposite cytosine or thymidine
-
-
-
additional information
?
-
Escherichia coli MV1161
-
repairs 5-formyluracil, a major thymine lesion produced by reactive oxygen radicals and photosensitized oxidation, 5-hydroxymethyluracil, another major thymine methyl oxidation product, is not a substrate
-
-
-
additional information
?
-
Escherichia coli MV1161
-
also capable to relase xanthine and oxanine, guanine lesions induced by nitrosation
-
-
-
additional information
?
-
Escherichia coli JM105
-
enzyme cleaves pBR322 and pAlk10 plasmids, excises adducts formed by chloroacetaldehyde, acrolein and croton aldehyde but is not able to excise malionaldehyde and bulky p-benzochinone adducts
-
-
-
additional information
?
-
Escherichia coli BW 9062
-
enzyme releases N1-(carboxyethyl)adenine and N7-(carboxymethyl)guanine from DNA treated with beta-propiolactone, but does not release the aflatoxin B-1 adduct at N-7 of guanine, release of 3-methyladenine from single-stranded DNA is 9% of the rate from double-stranded DNA, enzyme will not release 2,6-diamino-4-hydroxy-5-(N-methylformamido)pyrimidine, the alkali-induced derivative of 7-methylguanine, in which the imidazole ring is opened
-
-
-
additional information
?
-
Escherichia coli AB1157
-
repairs 5-formyluracil, a major thymine lesion produced by reactive oxygen radicals and photosensitized oxidation, 5-hydroxymethyluracil, another major thymine methyl oxidation product, is not a substrate
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
cellular repair of alkylated DNA base modifications
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
DNA repair enzyme, inducible base excision repair pathway of DNA alkylation damage
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
P04395
DNA repair enzyme, inducible base excision repair pathway of DNA alkylation damage
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
induced by cell exposure to sublethal doses of alkylating agents, important role in the excision of base damage from single-stranded regions transiently formed in DNA during transcription and replication
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
short-patch base excision repair pathway, repair of alkylation and oxidative DNA damage
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
binds mismatch base-pairs in DNA
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
-
inducible pathway for repair of DNA damaged by simple alkylating agents such as methylmethanesulfonate and N-methyl-N'-nitro-N-nitrosoguanidine
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
Escherichia coli MV1161
-
inducible pathway for repair of DNA damaged by simple alkylating agents such as methylmethanesulfonate and N-methyl-N'-nitro-N-nitrosoguanidine
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
Escherichia coli MV1161
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
Escherichia coli JM105
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
Escherichia coli BW 9062
-
-
-
-
?
alkylated DNA + H2O
3-methyladenine + 7-methylguanine + O2-methylthymine + O2-methylcytosine + DNA
show the reaction diagram
Escherichia coli AB1157
-
DNA repair enzyme, inducible base excision repair pathway of DNA alkylation damage
-
-
?
additional information
?
-
-
AlkD is involved exclusively in the repair of alkylation damage
-
-
-
additional information
?
-
-
enzyme is involved in DNA excision repair
-
-
-
additional information
?
-
-
the enzyme is induced in response to DNA alkylation, and it protects cells from alkylated nucleobases by catalyzing their excision
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
additional information
-
no absolute cofactor requirement
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ca2+
-
2 mM CaCl2, enzyme activity 103%
Cd2+
P29372
inhibitory above 0.05 mM. Reduced catalytic activity in presence of Zn2+ is not due to altered binding of substrate
K+
-
optimal activity at pH 7.5 and 100 mM KCl
Mg2+
-
stimulated by 1 mM
Mg2+
-
stimulated by 1.5 mM MgCl2
Mg2+
-
no obligatory requirement, but addition of 1.5 mM MgCl2 causes stimulation in the catalytic activity 2fold
Mg2+
-
2 mM MgSO4, enzyme activity 112%
Mg2+
-
at low concentration, Mg2+ stimulates enzyme activity with substrate hypoxanthine, but not with ethenoadenine. At high concentrations, inhibitory
Mn2+
-
2 mM MnCl2, enzyme activity 102%
Ni2+
P29372
inhibitory above 0.05 mM, but to lesser extent than Cd2+ or Zn2+. Reduced catalytic activity in presence of Zn2+ is not due to altered binding of substrate
Zn2+
-
0.1 mM ZnSO4, enzyme activity 114%
Zn2+
P29372
enzyme active site has a potential binding site for Zn2+. Reduced catalytic activity in presence of Zn2+ is not due to altered binding of substrate
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2'-deoxy-2'-fluoro-7-methylguanosine
-
-
-
abasic-containing DNA
-
potent inhibitor
-
apurinic-DNA
-
-
-
DNA containing 1-azaribose
-
-
-
Double-stranded DNA
-
-
N-ethylmaleimide
-
1 mM, 24% inhibition
Ni2+
-
0.001 mM
p-hydroxymercuribenzoate
-
-
p-mercuribenzoate
-
-
Pyrrolidine-containing oligonucleotide
-
-
-
ZnSO4
-
2 mM, enzyme activity 23%
Mg2+
-
significant inhibition depending on its concentration, but independent of substrate type. Mg2+ at high but physiologic concentrations decreases the amount of active enzyme. Inhibition affects the Km value, but not Vmax, and is due to inhibition in substrate binding and can be reversed by EDTA. At low concentration, Mg2+ stimulates enzyme activity with substrate hypoxanthine, but not with ethenoadenine
additional information
-
prior complex formation between mismatch repair protein MutS and a heteroduplex containing an 5-formyluracil-guanine mispair inhibits the activity of AlkA to 5-formyluracil
-
additional information
-
resistant to product inhibition by free 3-methyladenine
-
additional information
-
resistant to product inhibition by free 3-methyladenine
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
3-Methyladenine
-
5 mM, enzyme activity 110%
3-Methylguanine
-
5 mM, enzyme activity 120%
N-methyl-N'-nitro-N-nitrosoguanidine
-
-
N-methyl-N'-nitro-N-nitrosoguanidine
-
MNNG, good inducer of the enzyme, methylmethanesulfonate is less effective and 4-nitroquinoline-1-oxide has no effect
spermidine
-
stimulates activity
spermine
-
stimulates activity
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.000024
-
1,N6-ethenoadenine residues in alkylated DNA
-
pH 7.5, 37C, excision of 1, N6-ethenoadenine
-
0.00004
0.001
1,N6-ethenoadenine residues in alkylated DNA
-
pH 8.3, 37C, release of 1,N6-ethenoadenine, wild type enzyme
-
0.00006
0.0013
1,N6-ethenoadenine residues in alkylated DNA
-
pH 8.3, 37C, release of 1,N6-ethenoadenine, truncated mutant N-DELTA100C-DELTA18
-
0.0000092
-
3-methyladenine residues in alkylated DNA
-
pH 8.0, 37C, release of 3-methyladenine
-
0.000011
-
7-methylguanine residues in alkylated DNA
-
pH 8.0, 37C, release of 7-methylguanine
-
0.00000014
-
Alkylated DNA
-
pH 7.5, 37C, excision of 5-hydroxymethyluracil, denatured DNA
0.0000044
-
Alkylated DNA
-
pH 7.5, 37C, excision of 5-hydroxymethyluracil
0.0000108
-
Alkylated DNA
-
pH 7.5, 37C, excision of 3-methyladenine
0.000021
-
Alkylated DNA
-
pH 8.0, 37C, excision of 5-formyluracil
0.00003
-
Alkylated DNA
-
pH 8.0, 37C, excision of 7-methylguanine
0.000031
-
alkylated DNA, treated with N-methyl-N'-nitro-N-nitrosoguanidine
-
pH 7.5, 37C, excision of 7-methylguanine
-
0.000053
-
alkylated DNA, treated with N-methyl-N'-nitro-N-nitrosoguanidine
-
pH 7.5, 37C, excision of xanthine
-
0.0000354
-
duplex oligonucleotide substrate containing ethenoadenine and hypoxanthine
-
37C, pH 7.9
-
0.0000448
-
duplex oligonucleotide substrate containing ethenoadenine and hypoxanthine
-
37C, pH 7.9, presence of 250 mM Mg2+
-
0.0000548
-
duplex oligonucleotide substrate containing ethenoadenine and hypoxanthine
-
37C, pH 7.9, presence of 500 mM Mg2+
-
0.042
-
duplex oligonucleotide substrate containing ethenoadenine and hypoxanthine
-
at 37C, in 50 mM Na-MES buffer (pH 6.1)
-
0.0000053
-
hypoxanthine
-
at 37C
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.000833
-
1,N6-ethenoadenine residues in alkylated DNA
-
pH 7.5, 37C, excision of1, N6-ethenoadenine
-
0.01
-
1,N6-ethenoadenine residues in alkylated DNA
-
pH 8.3, 37C, release of 1,N6-ethenoadenine, truncated mutant N-DELTA100C-DELTA18
-
0.0167
-
1,N6-ethenoadenine residues in alkylated DNA
-
pH 8.3, 37C, release of 1,N6-ethenoadenine, wild-type
-
0.00125
-
duplex oligonucleotide substrate containing ethenoadenine and hypoxanthine
-
at 37C, in 50 mM Na-MES buffer (pH 6.1)
-
0.0033
-
hypoxanthine
-
at 37C
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
30
-
duplex oligonucleotide substrate containing ethenoadenine and hypoxanthine
-
at 37C, in 50 mM Na-MES buffer (pH 6.1)
0
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.00000008
-
-
mutant Asn-152, expressed in Escherichia coli
0.000005
-
-
mutant Asn-252, expressed in Escherichia coli
0.000007
-
-
mutant Asn-255 and Asn-231, expressed in Escherichia coli
0.000009
-
-
mutant Asn-259, expressed in Escherichia coli
0.00001
-
-
mutant Asn-260, expressed in Escherichia coli
0.000013
-
-
mutant Asn-112, expressed in Escherichia coli
0.000014
-
-
wild-type Ndelta100Cdelta18, expressed in Escherichia coli
0.000018
-
-
mutant Asn-310, expressed in Escherichia coli
0.000375
-
-
-
0.0247
-
-
-
additional information
-
-
specific activity 1100 pmol of 3-methyladenine released/mg of protein, 6.7 mg protein/4 ml
additional information
-
-
the crude extract shows a specific activity of 80 IU/mg, the 125fold purified enzyme displays a specific activity of 10000 IU/mg, one international unit (IU) is defined as the activity, which cleaves 50% of 0.5 pmole of double stranded DNA oligo substrate containing a hypoxanthine lesion in 10 min at 37C
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.5
-
-
-
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.3
7.4
-
isoelectric focusing
9.65
-
-
isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
normal, malignant and immortalized breast cells
Manually annotated by BRENDA team
-
immortalized, non-transformed breast epithelial cell line
Manually annotated by BRENDA team
-
thymic carcinoma
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Schizosaccharomyces pombe (strain 972 / ATCC 24843)
Schizosaccharomyces pombe (strain 972 / ATCC 24843)
Schizosaccharomyces pombe (strain 972 / ATCC 24843)
Schizosaccharomyces pombe (strain 972 / ATCC 24843)
Schizosaccharomyces pombe (strain 972 / ATCC 24843)
Schizosaccharomyces pombe (strain 972 / ATCC 24843)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
22000
-
-
gel filtration, Svedberg equation
27000
-
-
gel filtration
31000
-
-
SDS-PAGE
32000
-
-
SDS-PAGE
33000
-
-
SDS-PAGE
34250
-
-
predicted from nucleotide sequence
40000
-
-
recombinant protein, expressed in Escherichia coli, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 25000, AlkD, SDS-PAGE
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
at 1.8 A resolution. Structure contains two sodium ions in octahedral coordination
-
sitting drop vapour diffusion method, with 85 mM HEPES (pH 7.5), 15% (w/v) PEG 4000, and 17% (v/v) glycerol, at 21C
-
2.5 A crystal structure colmplexed to DNA, crystals belong to space group P3(1)21, cell dimensions a = B = 82.4 A and c = 199.7 A
-
AlkA in complex with undamaged DNA, sitting drop vapor diffusion method, using 25-29% (w/v) polyethylene glycol 3350, 100 mM bis-Tris, pH 6.0-6.6, 200 mM Li2S04, and 3% (w/v) 6-aminocaproic acid, at 25C
-
crystal structure of AlkA solved with the multiple isomorphous replacement method, crystals grown with sitting-drop vapor-diffusion technique, space group P2(1), cell dimensions a = 58.61 A, b = 76.93 A, c = 62.27 A
-
hanging drop vapor difusion crystallization, space group P2(1), unit-cell parameters are a = 58.17, b = 76.27, c = 61.69, alpha = gamma = 90, beta = 109.98
-
sitting drop method of vapour diffusion, monoclinic space group C2, a = 58.6 A, b = 76.8 A, c = 62.2 A
-
AAG complexed to DNA
-
expression in Saccharomyces cerevisiae
-
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
45
-
-
activity reduced by 50%
48
-
-
relatively heat resistant
48
-
-
50% inactivation requires 65 min
50
-
-
65 min time for 50% inactivation
50
-
-
3.5 min, 50% inactivation
50
-
-
the purified full-length MPG is stable at high temperature, at 50 C 30 min of incubation inactivate the protein by 30%
60
-
-
heat stable, retains 40% of its original activity after 2 min incubation
90
-
-
60% enzyme activity retains after incubation for 1 h
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
very unstable
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-70C, purified enzyme remains stable for more than 4 years when stored in 20% glycerol,
-
-20C, stored in 50% glycerol, undergoes complete inactivation in 3-4 weeks
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Ni-NTA column chromatography, heparin affinity column chromatography, and gel filtration
-
Mono Q column chromatography
-
purified from an overproducing strain
-
purified from overproducing strains DH1 and HB101, harboring plasmid pYN3070
-
purified from strain JM105 harboring pAlk10 plasmid
-
recombinant protein, purified from Escherichia coli
-
SP Sepharose column chromatography
-
recombinant protein, expressed from baculovirus
-
recombinant protein, expressed in Escherichia coli
-
recombinant protein, purified from Escherichia coli
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
gene afalkA, AF2117 ORF cloned and overexpressed in Escherichia coli, gene function suppresses the alkylation sensitivity in Escherichia coli tag alkA double mutants
-
expressed in Escherichia coli HMS174 cells
-
expression in Escherichia coli BL21, AlkD
-
alkAg+ gene cloned and sequenced
-
expressed in Escherichia coli
-
construction of recombinant GST-MPG fusion proteins
-
expressed in Escherichia coli BL21(DE3) cells
-
overexpressed in MDA-MB231 breast cancer cell line
-
cloned and expressed in Escherichia coli BL21(DE3)
-
MPG deletion mutant N-DELTA100C-DELTA18 overexpressed in Escherichia coli
-
truncated MPG polypeptides expressed in Escherichia coli using vector pRSET, full length MPG protein cloned in eukaryotic baculovirus expression vector pVL1393 and host Sf9 insect cells used for expression
-
expression by adenoviral system and targeting of enzyme to mitochondria or nucleus of primary astrocytes
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
D113N
-
the mutation results in a 100fold decrease in the single-turnover rate of 7-methylguanine excision relative to the wild type enzyme
R148A
-
the mutation results in a 100fold decrease in the single-turnover rate of 7-methylguanine excision relative to the wild type enzyme
D238N
-
site-directed mutagenesis
W218A
-
site-directed mutagenesis
N169A
-
ratio of 3-methyladenine/7-methylguanine is 29:1 for wild-type enzyme and 26:1 for mutant enzyme N169A, 30fold lower activity than wild-type enzyme. Expression of the N169A in Saccharomyces cerevisiae during methyl methanesulfonate exposure results in greater sensitivity, greater mutation induction following exposure to methyl methanesulfonate and more strand breaks in vivo
N169D
-
ratio of 3-methyladenine/7-methylguanine is 29:1 for wild-type enzyme, 100fold lower production of 3-methyladeine and no production of 7-methylguanine is detected with mutant enzyme N169D. When expressed in Saccharomyces cerevisiae, the N169D variant provides better protection against methyl methanesulfonate toxicity than wild-type enzyme. Fewer strand breaks in vivo are seen in the presence of the N169D variant following exposure to methyl methanesulfonate
N169S
-
ratio of 3-methyladenine/7-methylguanine is 29:1 for wild-type enzyme and 24:1 for mutant enzyme N169S, 30fold lower activity than wild-type enzyme. Expression of the N169S in Saccharomyces cerevisiae during methyl methanesulfonate exposure results in greater sensitivity, greater mutation induction following exposure to methyl methanesulfonate and more strand breaks in vivo
D112N
-
site-directed mutagenesis
D152N
-
site-directed mutagenesis
D252N
-
site-directed mutagenesis
D259N
-
site-directed mutagenesis
D260N
-
site-directed mutagenesis
D310N
-
site-directed mutagenesis
additional information
-
recombinant expression of enzyme and targeting to mitochondria or nucleus of primary astrocytes. Increasing enzyme activity significantly increases base excision repair kinetics in both the mitochondria and nuclei. Increased nuclear enzyme activity results in cell death in astrocyte cultures treated with methylnitrososurea
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
inhibition by Cd2+, Ni2+, or Zn2+ can be reversed by EDTA
P29372
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
medicine
-
sulfur mustard, or mustard gas, bis-(2-chloroethyl)sulfide is carcinogenic to humans, acutely toxic to the skin, repiratory tract and eyes and causes a delayed bone marrow depression, knowledge of the DNA damage caused by this agent and the cellular defenses against this damage are of fundamental importance
medicine
-
MPG overexpression is paradoxically associated with increased suspectibility to DNA damage, up-regulation of this gene may suggest a functional role in breast carcinogenesis, animal models using transgenic mice overexpressing the enzyme are being developed to determine the in vivo role of MPG in breast carcinogenesis
medicine
-
overexpression of MPG can result in increased kill of tumor cells using lower doses of harmful alkylating agents such as temozolomide and cross-linking agents such as cisplatin, use of lower doses of chemotherapeutic agents, which will decrease the emergence of drug-resistant tumor cells and decrease the need fro stem-cell support, leading to an increased patient response and a better quality of life for the patient
medicine
-
recombinant expression of enzyme and targeting to mitochondria or nucleus of primary astrocytes. Increasing enzyme activity significantly increases base excision repair kinetics in both the mitochondria and nuclei. Increased nuclear enzyme activity results in cell death in astrocyte cultures treated with methylnitrososurea