| Description | Fpg (also known as formamidopyrimidine [fapy]-DNA glycosylase, Mut M, FAPY DNA Glycosylase, and 8-oxoguanine DNA glycosylase) has both N-glycosylase and AP-lyase activities. The N-glycosylase activity releases damaged purines from double stranded DNA, generating an apurinic (AP site). The AP-lyase activity cleaves both 3´ and 5´ to the AP site thereby removing the AP site and producing a 1 base gap in the DNA and 3′ and 5′ phosphate termini. Bases recognized and removed by Fpg include 7, 8-dihydro-8-oxoguanine (8-oxoguanine), 8-oxoadenine, fapy-guanine, methy-fapy-guanine, fapy-adenine, aflatoxin B1-fapy-guanine, 5-hydroxy-cytosine and 5-hydroxy-uracil (1,2). |
| Components | - E. coli fpg: 8,000 U/ml in 20 mM Tris-HCl, pH 8.0 @ 25°C, 50 mM NaCl, 0.5 mM EDTA, 200 μg/ml BSA, 50% Glycerol
- 10X Reaction Buffer: 100 mM Bis-Tris-Propane, 100 mM MgCl2, 10 mM DTT, pH 7.0 @ 25°C
|
| References | Tchou, J. et al. (1994) Substrate specificity of Fpg protein. J. Biol. Chem., 269, 15318-15324.Hatahet, Z. et al. (1994) New substrates for old enzymes. J. Biol. Chem., 269, 18814-18820.Boiteux, S., O’Connor, T. and Laval, J. (1987) Formamidopyrimidine-DNA glycosylase of Escherichia coli: cloning and sequencing of the fpg structural gene and overproduction of the protein. EMBO J., 5, 3177-3183.Singh, N., McCoy, M., Tice, R. and Schneider, L. (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp. Cell Res., 175, 184-191.Collins, A., Duthie, S. and Dobson, V. (1993) Direct enzymatic detection of endogenous oxidative base damage in human lymphocyte DNA. Carcinogenesis, 14, 1733-1735.Collins, A., Dusinska, M., Gedik, C. and Stetina, R. (1996) Oxidative damage to DNA: do we have a reliable biomarker?. Environ. Health Perspect., 104, 465-469.Pflaum, M., Will, O., Mahler, H.-C. and Epe, B. (1998) DNA oxidation products determined with repair endonucleases in mammalian cells: types, basal levels and influence of cell proliferation. Free Rad. Res., 29, 585-594.Hartwig, A., Dally, H. and Schlepegrell, R. (1996) Sensitive analysis of oxidative DNA damage in mammalian cells: use of the bacterial Fpg protein in combination with alkaline unwinding. Toxicol. Lett., 88, 85-90.Czene, S. and Harms-Ringdahl, M. (1995) Detection of single strand breaks and formamidopyrimidine-DNA glycosylase-sensitive sites in DNA of cultured human fibroblasts. Mutat. Res., 336, 235-242. |
