title: Mass Spectrometry-Based Tools to Characterize DNA-Protein Cross-Linking by Bis-Electrophiles.
abstract: DNA-protein cross-links (DPCs) are unusually bulky DNA adducts that form in cells as a result of exposure to endogenous and exogenous agents including reactive oxygen species, ultraviolet light, ionizing radiation, environmental agents (e.g. transition metals, formaldehyde, 1,2-dibromoethane, 1,3-butadiene) and common chemotherapeutic agents. Covalent DPCs are cytotoxic and mutagenic due to their ability to interfere with faithful DNA replication and to prevent accurate gene expression. Key to our understanding of the biological significance of DPC formation is identifying the proteins most susceptible to forming these unusually bulky and complex lesions and quantifying the extent of DNA-protein cross-linking in cells and tissues. Recent advances in bottom-up mass spectrometry-based proteomics have allowed for an unbiased assessment of the whole protein DPC adductome after in vitro and in vivo exposures to cross-linking agents. This MiniReview summarizes current and emerging methods for DPC isolation and analysis by mass spectrometry-based proteomics. We also highlight several examples of successful applications of these novel methodologies to studies of DPC lesions induced by bis-electrophiles such as formaldehyde, 1,2,3,4-diepoxybutane, nitrogen mustards and cisplatin.
authors.affiliation: Department of Medicinal Chemistry, Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA.
authors.forename: Arnold
authors.initials: A
authors.lastname: Groehler
authors.affiliation: Department of Medicinal Chemistry, Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA.
authors.forename: Amanda
authors.initials: A
authors.lastname: Degner
authors.affiliation: Department of Medicinal Chemistry, Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA.
authors.forename: Natalia Y
authors.initials: NY
authors.lastname: Tretyakova
mesh_terms: D002945:Cisplatin / Q000633:toxicity; D003432:Cross-Linking Reagents / Q000633:toxicity*; D004247:DNA / Q000737:chemistry; D018736:DNA Adducts / Q000032:analysis* / Q000737:chemistry / Q000633:toxicity; D004249:DNA Damage; D004781:Environmental Exposure / Q000009:adverse effects*; D004852:Epoxy Compounds / Q000633:toxicity; D005557:Formaldehyde / Q000633:toxicity; D013058:Mass Spectrometry / Q000379:methods*; D009152:Mutagenicity Tests / Q000379:methods; D009153:Mutagens / Q000633:toxicity; D009588:Nitrogen Mustard Compounds / Q000633:toxicity; D011506:Proteins / Q000737:chemistry; D040901:Proteomics / Q000379:methods
chemical_list: D003432:Cross-Linking Reagents; D018736:DNA Adducts; D004852:Epoxy Compounds; D009153:Mutagens; D009588:Nitrogen Mustard Compounds; D011506:Proteins; D005557:Formaldehyde; C007366:diepoxybutane; D004247:DNA; D002945:Cisplatin