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Bifunctional DNA glycosylase with DNA N-glycosylase and AP lyase activities
N-glycosylase activity releases 8-oxo-7,8-dihydroguanine (8-oxoG), generating an AP site. The AP-lyase activity cleaves 3′ to the AP site leaving a 5′ phosphate and a 3′-phospho-α, β-unsaturated aldehyde.
Thermostable OGG is an archaeal 8-oxoguanine (8-oxoG) DNA glycosylase which acts both as a N-glycosylase and an AP-lyase. The N-glycosylase activity releases the damaged purine (8-oxoguanine) from double stranded DNA, generating an apurinic (AP) site. The AP-lyase activity cleaves 3′ to the AP site leaving a 5′ phosphate and a 3′-phospho-α, β-unsaturated aldehyde. Unlike some other DNA glycosylases, Thermostable OGG specifically recognizes and cleaves only 8-oxoG and no other modified bases.
Figure 1: Unlike Fpg, Thermostable OGG specifically recognizes and cleaves only 8-oxoguanine and no other modified bases.
The ability to excise modified bases was queried by incubating Thermostable OGG (8 U) or Fpg (8 U) with fluorescently labeled dsDNA containing modified bases (either 8-oxoadenine, 5-hydroxycytosine, thymine glycol, 5-hydroxyuridine or 8-oxoguanine). Samples were analyzed by capillary electrophoresis and cleavage products are evidenced by the appearance of a cleaved product peak. Thermostable OGG only cleaves the 8-oxoguanine sample (top panel) while Fpg is active on other substrates including , 5-hydroxycytosine, thymine glycol and 5-hydroxyuridine (bottom panel).
Highlights
Isolated from a recombinant source
Supplied with 10X Reaction Buffer
Product Source
An E. coli strain that carries a cloned thermostable archael oxoguanine glycosylase gene.
This product is related to the following categories:
One unit is defined as the amount of enzyme required to cleave 10 pmol of a 60-mer fluorescently labeled oligonucleotide duplex containing a single 8-oxoguanine base paired with a cytosine in a total reaction volume of 10 μl in 1 hour at 65°C.
Reaction Conditions
1X rCutSmart™ Buffer
Incubate at 65°C
1X rCutSmart™ Buffer
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
100 µg/ml Recombinant Albumin
(pH 7.9 @ 25°C)
Storage Buffer
10 mM Tris-HCl
500 mM NaCl
0.1 mM EDTA
1 mM DTT
200 µg/ml Recombinant Albumin
50% Glycerol
pH 7.4 @ 25°C
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