Department of Medicinal Chemistry, Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA.
Basic Clin Pharmacol Toxicol. 2017 Sep;121 Suppl 3(Suppl 3):63-77. doi: 10.1111/bcpt.12751. Epub 2017 Mar 14.
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.
DNA-蛋白质交联物（DPCs）是一种异常大体积的 DNA 加合物，由于暴露于内源性和外源性物质（如活性氧、紫外线、电离辐射、环境物质（如过渡金属、甲醛、1,2-二溴乙烷、1,3-丁二烯）和常见的化疗药物）而在细胞中形成。由于其能够干扰忠实的 DNA 复制并防止准确的基因表达，因此共价 DPCs 具有细胞毒性和致突变性。了解 DPC 形成的生物学意义的关键是确定最容易形成这些异常大体积和复杂损伤的蛋白质，并定量测定细胞和组织中的 DNA-蛋白质交联程度。基于质谱的蛋白质组学的最新进展允许在体外和体内暴露于交联剂后，对整个蛋白质 DPC 加合物组进行无偏评估。本综述总结了目前和新兴的基于质谱的蛋白质组学 DPC 分离和分析方法。我们还强调了这些新方法在研究甲醛、1,2,3,4-二环氧丁烷、氮芥和顺铂等双电子亲和试剂诱导的 DPC 损伤方面的几个成功应用实例。
