Synthesis of 2'-deoxyoxanosine from 2'-deoxyguanosine, conversion to its phosphoramidite, and incorporation into oxanine-containing oligodeoxynucleotides.
Oxanine (Oxa, O) is one of the damaged bases produced from guanine (G) through nitrosative deamination induced by nitric oxide (NO) or nitrous acid (HNO(2)). Large-scale preparation of Oxa-containing oligodeoxynucleotide (Oxa-ODN) with the desired base sequence is a prerequisite for exploring detailed properties of Oxa in DNA. This can be accomplished by incubation of G nucleosides with NaNO(2) in acetic acid buffer (pH 3.5) to produce Oxa nucleosides (e.g., 2'-deoxyoxanosine or dOxo), conversion of dOxo to DMT-dOxo-amidite by tritylation and conventional phosphoramidation, and subsequent synthesis of Oxa-ODN. The presence of Oxa in the synthetic ODN is confirmed by enzymatic digestion. Oxa-ODN is useful for analyzing the biochemical and biophysical properties of Oxa in DNA, which is believed to be involved in NO-induced genotoxicity and cytotoxicity. In addition, since Oxa possesses the carbodiimide-activated carboxylate function (O-acylisourea structure), Oxa-ODN can be used as a functional DNA oligomer that makes covalent cross-linkages with amine or amine-containing biomolecules and amine-modified solid surfaces.
Korea University, Jochiwon, Chungnam, Korea.
Seung Pil
SP
Pack
Keisuke
K
Makino
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003849:Deoxyguanosine / Q000737:chemistry*; D003853:Deoxyribonucleosides / Q000138:chemical synthesis / Q000737:chemistry; D008956:Models, Chemical; D015394:Molecular Structure; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis* / Q000737:chemistry; D009943:Organophosphorus Compounds / Q000737:chemistry*; D011684:Purine Nucleosides / Q000737:chemistry
D003853:Deoxyribonucleosides; D009838:Oligodeoxyribonucleotides; D009943:Organophosphorus Compounds; D011684:Purine Nucleosides; C468766:oxanine; C434331:phosphoramidite; C042917:2'-deoxyoxanosine; D003849:Deoxyguanosine