title: Nuclease P1 digestion combined with tandem mass spectrometry for the structure determination of DNA photoproducts.
abstract: 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.
authors.affiliation: Department of Chemistry, Washington University, St. Louis, Missouri 63130, USA.
authors.forename: Y
authors.initials: Y
authors.lastname: Wang
authors.forename: J S
authors.initials: JS
authors.lastname: Taylor
authors.forename: M L
authors.initials: ML
authors.lastname: Gross
mesh_terms: D018360:Crystallography, X-Ray; D004247:DNA / Q000737:chemistry* / Q000528:radiation effects; D053938:DNA Fragmentation; D006868:Hydrolysis; D013058:Mass Spectrometry; D009841:Oligonucleotides / Q000737:chemistry / Q000528:radiation effects; D010777:Photochemistry; D010782:Photolysis; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014466:Ultraviolet Rays
chemical_list: D009841:Oligonucleotides; D004247:DNA; D015719:Single-Strand Specific DNA and RNA Endonucleases