HPLC-32P-postlabelling analysis of 1,N6-ethenodeoxyadenosine and 3,N4-ethenodeoxycytidine.
A 32P-postlabelling procedure coupled with HPLC has been developed to detect and measure the cyclic nucleic acid adducts 1,N6-ethenodeoxyadenosine and 3,N4-ethenodeoxycytidine in DNA. Chloroacetaldehyde-modified DNA containing these adducts was enzymatically digested to 3'-monophosphates and adducts were separated by ion-pair reverse-phase HPLC on PL RP-S prior to 32P-postlabelling with carrier free [gamma-32P]ATP. Following 3'-dephosphorylation with nuclease P1 the resulting [5'-32P]monophosphate adducts were finally resolved by HPLC on PL RP-S and assayed by liquid scintillation counting.
Watson
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WP
Shell Research Ltd, Sittingbourne Research Centre, Kent, UK.
Crane
A E
AE
D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000378:metabolism; D003839:Deoxyadenosines / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism; D003845:Deoxycytosine Nucleotides / Q000032:analysis* / Q000378:metabolism; D010761:Phosphorus Radioisotopes; D013950:Thymus Gland / Q000032:analysis
D003839:Deoxyadenosines; D003845:Deoxycytosine Nucleotides; D010761:Phosphorus Radioisotopes; D003841:Deoxycytidine; C032221:1,N(6)-ethenodeoxyadenosine; C032222:3,N(4)-ethenodeoxycytidine; D004247:DNA