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Information on EC 2.1.1.63 - methylated-DNA-[protein]-cysteine S-methyltransferase and Organism(s) Mycobacterium tuberculosis and UniProt Accession P9WJW5

for references in articles please use BRENDA:EC2.1.1.63
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EC Tree
IUBMB Comments
This protein is involved in the repair of methylated DNA. Unlike EC 3.2.2.20, DNA-3-methyladenine glycosidase I and EC 3.2.2.21, DNA-3-methyladenine glycosidase II, which remove the methylated base leaving an apurinic/apyrimidinic site, this enzyme transfers the methyl group from the methylated DNA to an internal cysteine residue, leaving an intact nucleotide. Since the methyl transfer is irreversible, the enzyme can only catalyse a single turnover.
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This record set is specific for:
Mycobacterium tuberculosis
UNIPROT: P9WJW5
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Word Map
The taxonomic range for the selected organisms is: Mycobacterium tuberculosis
The enzyme appears in selected viruses and cellular organisms
Synonyms
o6-methylguanine-dna methyltransferase, atase, o6-alkylguanine-dna alkyltransferase, ada protein, o6-methylguanine dna methyltransferase, o6-methylguanine-dna-methyltransferase, o6-methylguanine methyltransferase, o6-alkylguanine dna alkyltransferase, o6-mgmt, methylguanine dna methyltransferase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
O6-methylguanine methyltransferase
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O6-methylguanine-DNA methyltransferase
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Ada protein
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MGMT
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O6-methylguanine-DNA methyltransferase
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O6-MGMT
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
methyl group transfer
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SYSTEMATIC NAME
IUBMB Comments
DNA-6-O-methylguanine/DNA-4-O-methylthymine:[protein]-L-cysteine S-methyltransferase
This protein is involved in the repair of methylated DNA. Unlike EC 3.2.2.20, DNA-3-methyladenine glycosidase I and EC 3.2.2.21, DNA-3-methyladenine glycosidase II, which remove the methylated base leaving an apurinic/apyrimidinic site, this enzyme transfers the methyl group from the methylated DNA to an internal cysteine residue, leaving an intact nucleotide. Since the methyl transfer is irreversible, the enzyme can only catalyse a single turnover.
CAS REGISTRY NUMBER
COMMENTARY hide
77271-19-3
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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 6-O-methylguanine) + protein L-cysteine
DNA (without 6-O-methylguanine) + protein S-methyl-L-cysteine
show the reaction diagram
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
DNA (containing 6-O-methylguanine) + protein L-cysteine
DNA (without 6-O-methylguanine) + protein S-methyl-L-cysteine
show the reaction diagram
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
17844
x * 17844, calculated from amino acid sequence
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 17844, calculated from amino acid sequence
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
hanging drop vapor diffusion method, for wild type, enzyme using 0.2 M ammonium acetate, 22% (w/v) PEG 3350, and 0.1 M HEPES pH 7.5, while for mutant enzymes R37K and Y139F using 0.1 M HEPES pH 7.5, 4% polyethylene glycol 8000, and either 4% or 8% ethylene glycol, respectively
sitting drop vapor diffusion method, using 0.1 M HEPES (pH 7.5), and 6% (w/v) polyethylene glycol 8000
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
R37E
the mutant exhibits a 5fold lower affinity for the methylated duplex DNA compared to the wild type enzyme
R37K
the variant performs a sub-optimal alkylated-DNA repair in vitro compared to the wild type enzyme
T15S
the mutant exhibits a 2fold-lower affinity for double stranded methylated DNA compared to the wild type enzyme
Y139F
the mutant exhibits a 10fold lower affinity for the methylated duplex DNA compared to the wild type enzyme
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
HiTrapQ column chromatography, MonoQ column chromatography, HiTrap heparin column chromatography, and Superdex 200 gel filtration
Superdex 200 gel filtration
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Miggiano, R.; Perugino, G.; Ciaramella, M.; Serpe, M.; Rejman, D.; Pav, O.; Pohl, R.; Garavaglia, S.; Lahiri, S.; Rizzi, M.; Rossi, F.
Crystal structure of Mycobacterium tuberculosis O6-methylguanine-DNA methyltransferase protein clusters assembled onto damaged DNA
Biochem. J.
473
123-133
2016
Mycobacterium tuberculosis (P9WJW5), Mycobacterium tuberculosis
Manually annotated by BRENDA team
Miggiano, R.; Casazza, V.; Garavaglia, S.; Ciaramella, M.; Perugino, G.; Rizzi, M.; Rossi, F.
Biochemical and structural studies of the Mycobacterium tuberculosis O6-methylguanine methyltransferase and mutated variants
J. Bacteriol.
195
2728-2736
2013
Mycobacterium tuberculosis (P9WJW5), Mycobacterium tuberculosis
Manually annotated by BRENDA team