title: Sequence selective tagging of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) using PNAs.
abstract: 8-Oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) is a commonly formed DNA lesion that is useful as a biomarker for oxidative stress. Methods for detecting 8-oxodGuo at specific positions within DNA could be useful for correlating DNA damage with mutational hotspots and repair enzyme accessibility. We describe a method for covalently linking ('tagging') peptide nucleic acids (PNAs) containing terminal nucleophiles under oxidative conditions to 8-oxodGuo at specific sites within DNA. Several nucleophiles were examined and the ε-amine of lysine was selected for further studies. As little as 10 fmol of 8-oxodGuo were detected by gel shift using (32)P-labeled target DNA and no tagging of dG at the same site or 8-oxodGuo at a distal site was detected when potassium ferricyanide was used as oxidant in substrates as long as 221 bp.
authors.affiliation: Johns Hopkins University, Department of Chemistry, Baltimore, CA 21218, United States; Kongju National University, Department of Chemistry, 182, Shinkwan-dong, Kongju, Chungnam 314-701, Republic of Korea.
authors.forename: In Seok
authors.initials: IS
authors.lastname: Hong
authors.affiliation: Johns Hopkins University, Department of Chemistry, Baltimore, CA 21218, United States. Electronic address: mgreenberg@jhu.edu.
authors.forename: Marc M
authors.initials: MM
authors.lastname: Greenberg
mesh_terms: D000080242:8-Hydroxy-2'-Deoxyguanosine; D001483:Base Sequence; D004247:DNA / Q000737:chemistry; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000032:analysis / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D008969:Molecular Sequence Data; D015394:Molecular Structure; D018384:Oxidative Stress; D020135:Peptide Nucleic Acids / Q000737:chemistry*; D010761:Phosphorus Radioisotopes; D013329:Structure-Activity Relationship
chemical_list: D020135:Peptide Nucleic Acids; D010761:Phosphorus Radioisotopes; D000080242:8-Hydroxy-2'-Deoxyguanosine; D004247:DNA; D003849:Deoxyguanosine
keywords: 8-OxodGuo; Biosensors; DNA damage; Oxidative stress; PNA