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E0526
Sigma
Endonuclease III from Escherichia coli
≥90% (SDS-PAGE), recombinant, expressed in Escherichia coli (overexpressed), buffered aqueous glycerol solution, ≥10,000 units/mg protein
| Synonym: | Nth protein, Endo III |
| MDL number: | MFCD02265700 |
Description
| Preparation Note | Purified in an iron bound form that confers the protein a light green color. |
| Unit Definition | One unit will cleave 50% of 0.5 pmol double strand oligonucleotide substrate containing 5,6-dihydrothymine in 15 min. at 30 °C. |
| Physical form | Solution in 20 mM Tris, pH 7.4, 100 mM NaCl, 1 mM DTT, 1 mM EDTA, and 50% glycerol. |
| Biochem/physiol Actions | Endo III, product of the nth gene, is an iron-sulfur containing DNA repair enzyme. It possesses DNA glycosylase activity with broad substrate specificity for mutated pyrimidine derivatives especially thymine glycol (Tg) but also for 5-hydroxycytosine and 5,6-dihydrothymine. In addition, it has an apurinic/apyrimidinic (AP) lyase activity which cleaves the DNA backbone by β elimination. Endo III protects E. coli cells from radiation damage caused by x-ray and UV irradiation and its activity overlaps the activity of endonuclease VIII. When overexpressed, it can also protect E.coli cells from alkylation defects caused by methylmethanesulphonate. |
Properties
| recombinant | expressed in Escherichia coli (overexpressed) |
| assay | ≥90% (SDS-PAGE) |
| form | buffered aqueous glycerol solution |
| mol wt | mol wt ≥23 kDa (211 amino acids) |
| shipped in | wet ice |
| storage temp. | −20°C |
Safety
| WGK Germany | 1 |
References
| reference | Asahara, H., et al., Purification and characterization of Escherichia coli endonuclease III from the cloned nth gene. Biochemistry 28, 4444, (1989) |
| Cunningham, R.P., Endonuclease III is an iron-sulfur protein. Biochemistry 28, 4450, (1989) | |
| D'Ham, C., et al., Excision of 5,6-dihydroxy-5,6-dihydrothymine, 5,6-dihydrothymine, and 5-hydroxycytosine from defined sequence oligonucleotides by Escherichia coli endonuclease III and Fpg proteins: kinetic and mechanistic aspects. Biochemistry 38, 3335, (1999) | |
| Mazumder, A., et al., Stereochemical studies of the β-elimination reactions at aldehydic abasic sites in DNA: endonuclease III from Escherichia coli, sodium hydroxide, and Lys-Trp-Lys. Biochemistry 30, 1119, (1991) | |
| Jiang, D., et al., Chemosensory and photosensory perception in purple photosynthetic bacteria utilize common signal transduction components. J. Bacteriol. 179, 3773, (1997) | |
| Serafini, D.M., Schellhorn, H.E., Endonuclease III and endonuclease IV protect Escherichia coli from the lethal and mutagenic effects of near-UV irradiation. Can. J. Microbiol. 45, 632, (1999) | |
| Eide, L., et al., Overexpression of endonuclease III protects Escherichia coli mutants defective in alkylation repair against lethal effects of methylmethanesulphonate. FEBS Lett. 491, 56, (2001) |
