E0651
Endonuclease VIII from Escherichia coli
≥90% (SDS-PAGE), recombinant, expressed in E. coli, buffered aqueous glycerol solution, ≥20,000 units/mg protein
Synonym(s):
Endo VIII, nei Protein
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About This Item
MDL number:
UNSPSC Code:
12352200
recombinant
expressed in E. coli
Assay
≥90% (SDS-PAGE)
form
buffered aqueous glycerol solution
specific activity
≥20,000 units/mg protein
mol wt
30 kDa (263 amino acid)
shipped in
wet ice
storage temp.
−20°C
Gene Information
Escherichia coli CFT073 ... nei(1035763)
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Biochem/physiol Actions
DNA repair protein, product of the nei gene, that recognizes and removes oxidized pyrimidines as thymine glycol (Tg) and 5,6-dihydrothymine from DNA. Endo VIII possesses both DNA glycosylase and apurinic/apyrimidinic (AP) lyase activities. Its oxidized-base substrate specificity overlaps the endonuclease III (endo III, nth) substrate specificity. This cross-reactivity is manifested in E.coli mutants. While E.coli nth and nei mutants are not sensitive or only slightly more sensitive to ionizing radiation and peroxides than wild type, the nei-nth double mutant is hypersensitive to oxidative stress. The mechanism used by endo VIII to cleave the DNA backbone (lyase activity) is β,γ-elimination, resembling the fpg protein. This differs from the mechanism of endo III, which cleaves the DNA backbone by β-elimination.
Physical form
Solution in 10% glycerol containing 20 mM Tris pH 7.4, 100 mM NaCl, 0.5 mM EDTA, 1mM DTT.
Other Notes
The amount of protein that cleaves 50% of 0.5 pmol double stranded DNA oligonucleotide substrate containing DHT in 10 min. at pH 7.6 at 30 °C.
Storage Class Code
10 - Combustible liquids
WGK
WGK 1
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Personal Protective Equipment
dust mask type N95 (US), Eyeshields, Gloves
Regulatory Information
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J Laval et al.
Mutation research, 402(1-2), 93-102 (1998-07-24)
As a consequence of oxidative stress, reactive oxygen species are generated in the cells. They interact with DNA and induce various modifications. Among them, oxidised purines (such as C8-oxoguanine and purines whose imidazole ring is opened), oxidised pyrimidines (such as
D Jiang et al.
Journal of bacteriology, 179(11), 3773-3782 (1997-06-01)
Escherichia coli possesses two DNA glycosylase/apurinic lyase activities with overlapping substrate specificities, endonuclease III and endonuclease VIII, that recognize and remove oxidized pyrimidines from DNA. Endonuclease III is encoded by the nth gene. Endonuclease VIII has now been purified to
C D'Ham et al.
Biochemistry, 38(11), 3335-3344 (1999-03-17)
Oligonucleotides that contain a single modified pyrimidine, i.e., thymine glycol (Tg), 5,6-dihydrothymine (DHT), and 5-hydroxycytosine (5-OHC) were synthesized in order to investigate the substrate specificity and the excision mechanism of two Escherichia coli repair enzymes: endonuclease III and formamidopyrimidine DNA
D Jiang et al.
The Journal of biological chemistry, 272(51), 32230-32239 (1998-01-24)
Escherichia coli endonuclease VIII (endo VIII) was identified as an enzyme that, like endonuclease III (endo III), removes radiolysis products of thymine including thymine glycol, dihydrothymine, beta-ureidoisobutyric acid, and urea from double-stranded plasmid or phage DNA and cleaves the DNA
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