Description | Fpg(alsoknownasformamidopyrimidine[fapy]-DNAglycosylase,MutM,FAPYDNAGlycosylase,and8-oxoguanineDNAglycosylase)hasbothN-glycosylaseandAP-lyaseactivities.TheN-glycosylaseactivityreleasesdamagedpurinesfromdoublestrandedDNA,generatinganapurinic(APsite).TheAP-lyaseactivitycleavesboth3´and5´totheAPsitetherebyremovingtheAPsiteandproducinga1basegapintheDNAand3′and5′phosphatetermini.BasesrecognizedandremovedbyFpginclude7,8-dihydro-8-oxoguanine(8-oxoguanine),8-oxoadenine,fapy-guanine,methy-fapy-guanine,fapy-adenine,aflatoxinB1-fapy-guanine,5-hydroxy-cytosineand5-hydroxy-uracil(1,2). |
Components | - E.colifpg:8,000U/mlin20mMTris-HCl,pH8.0@25°C,50mMNaCl,0.5mMEDTA,200μg/mlBSA,50%Glycerol
- 10XReactionBuffer:100mMBis-Tris-Propane,100mMMgCl2,10mMDTT,pH7.0@25°C
|
References | - Tchou,J.etal.(1994)SubstratespecificityofFpgprotein.J.Biol.Chem.,269,15318-15324.
- Hatahet,Z.etal.(1994)Newsubstratesforoldenzymes.J.Biol.Chem.,269,18814-18820.
- Boiteux,S.,O’Connor,T.andLaval,J.(1987)Formamidopyrimidine-DNAglycosylaseofEscherichiacoli:cloningandsequencingofthefpgstructuralgeneandoverproductionoftheprotein.EMBOJ.,5,3177-3183.
- Singh,N.,McCoy,M.,Tice,R.andSchneider,L.(1988)AsimpletechniqueforquantitationoflowlevelsofDNAdamageinindividualcells.Exp.CellRes.,175,184-191.
- Collins,A.,Duthie,S.andDobson,V.(1993)DirectenzymaticdetectionofendogenousoxidativebasedamageinhumanlymphocyteDNA.Carcinogenesis,14,1733-1735.
- Collins,A.,Dusinska,M.,Gedik,C.andStetina,R.(1996)OxidativedamagetoDNA:dowehaveareliablebioMarker?.Environ.HealthPerspect.,104,465-469.
- Pflaum,M.,Will,O.,Mahler,H.-C.andEpe,B.(1998)DNAoxidationproductsdeterminedwithrepairendonucleasesinmammaliancells:types,basallevelsandinfluenceofcellproliferation.FreeRad.Res.,29,585-594.
- Hartwig,A.,Dally,H.andSchlepegrell,R.(1996)SensitiveanalysisofoxidativeDNAdamageinmammaliancells:useofthebacterialFpgproteinincombinationwithalkalineunwinding.Toxicol.Lett.,88,85-90.
- Czene,S.andHarms-Ringdahl,M.(1995)Detectionofsinglestrandbreaksandformamidopyrimidine-DNAglycosylase-sensitivesitesinDNAofculturedhumanfibroblasts.Mutat.Res.,336,235-242.
|