Preparation of ethylenediaminephosphoramidates of nucleotides and derivatization with fluorescein isothiocyanate.
Fluorescence post-labeling of nucleotides is a potentially useful technique for the detection of trace amounts of damaged DNA (DNA adducts). Towards this goal, we have studied a derivatization procedure starting with the four major 5'-deoxynucleotides as model compounds. The 5'-phosphate group was first labeled with ethylenediamine via a phosphorimidazolide intermediate in methanol with an organic-soluble carbodiimide. The resulting ethylenediaminephosphoramidate products were reacted in turn with fluorescein isothiocyanate. The reaction sequence has been characterised at all stages by high-performance liquid chromatography.
al-Deen
A N
AN
Department of Biochemistry, Alexandria University, Egypt.
Cecchini
D C
DC
Abdel-Baky
S
S
Moneam
N M
NM
Giese
R W
RW
D055598:Chemical Phenomena; D002621:Chemistry; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000032:analysis / Q000302:isolation & purification; D004249:DNA Damage; D005029:Ethylenediamines / Q000032:analysis / Q000737:chemistry; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D007202:Indicators and Reagents; D009711:Nucleotides / Q000138:chemical synthesis*; D009943:Organophosphorus Compounds / Q000032:analysis / Q000737:chemistry; D013861:Thiocyanates
D005029:Ethylenediamines; D005452:Fluoresceins; D007202:Indicators and Reagents; D009711:Nucleotides; D009943:Organophosphorus Compounds; D013861:Thiocyanates; D004247:DNA; D016650:Fluorescein-5-isothiocyanate