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Nuclease P1 digestion combined with tandem mass spectrometry for the structure determination of DNA photoproducts

, : Nuclease P1 digestion combined with tandem mass spectrometry for the structure determination of DNA photoproducts. Chemical Research in Toxicology 12(11): 1077-1082, Nov

UV irradiation of oligodeoxynucleotides at 254 nm generates several different types of DNA photoproducts, such as cis-syn cyclobutane pyrimidine dimers, pyrimidine(6-4)pyrimidone photoproducts and their Dewar valence isomers, and thymine-adenine photoproducts (TA*). Nuclease P1 degrades the oligodeoxynucleotide photoproducts to small photoproduct-containing trinucleotides which are more amenable to tandem mass spectrometry (MS/MS) and HPLC. Product-ion mass spectra of these digestion products give characteristic fragmentations, allowing us to identify quickly the types of photomodifications. The results also show that mass spectrometry will be a tool for studying enzyme reaction mechanisms because it can determine rapidly and with high sensitivity the structures of the products that are generated.


PMID: 10563833

DOI: 10.1021/tx9900831

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