title: Kinetics and mechanism of DNA repair. Preparation, purification and some properties of caged dideoxynucleoside triphosphates.
abstract: Caged dideoxyribosylthymine triphosphate, dideoxyadenosine triphosphate and arabinosylcytosine triphosphate were prepared in high yield by reaction with 1-(2-nitrophenyl)diazoethane at pH 4 and room temperature for 24 h. Synthesis of caged alpha-32P-labelled dideoxyadenosine triphosphate (approx. 5000 Ci/mmol) in 85% yield was achieved by a modification of the method used for the synthesis of the unlabelled compounds. ATP was shown to be an excellent buffer in the synthesis of alpha-32P-labelled material, and in caged form to be an effective carrier in h.p.l.c. purification. Preparative h.p.l.c. was used to achieve purification of unlabelled caged compounds to greater than 98% purity and 32P-labelled material to 97% purity. Photolysis of unlabelled and 32P-labelled caged compounds by using XeF-excimer laser irradiation at 351 nm was characterized by using difference spectrophotometry and h.p.l.c. analysis. The stability of caged dideoxyadenosine [a-32P]triphosphate in the presence of cultured mammalian cells was evaluated; the adenosine derivative is essentially stable for 1 h.
authors.affiliation: Department of Biochemistry, University of Birmingham, U.K.
authors.forename: R A
authors.initials: RA
authors.lastname: Meldrum
authors.forename: S
authors.initials: S
authors.lastname: Shall
authors.forename: D R
authors.initials: DR
authors.lastname: Trentham
authors.forename: C W
authors.initials: CW
authors.lastname: Wharton
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D055598:Chemical Phenomena; D002621:Chemistry; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000528:radiation effects; D004249:DNA Damage; D004260:DNA Repair; D003838:Deoxyadenine Nucleotides; D015224:Dideoxynucleosides; D054306:Dideoxynucleotides; D000431:Ethanol / Q000031:analogs & derivatives; D007700:Kinetics; D051379:Mice; D009578:Nitrobenzenes; D010782:Photolysis; D013056:Spectrophotometry, Ultraviolet; D014466:Ultraviolet Rays
chemical_list: D003838:Deoxyadenine Nucleotides; D015224:Dideoxynucleosides; D054306:Dideoxynucleotides; D009578:Nitrobenzenes; C037773:2',3'-dideoxyadenosine triphosphate; C063593:1-(2-nitrophenyl)ethanol; D000431:Ethanol; D004247:DNA