ACTIVATION OF PROCARCINOGENS BY GLUTATHIONE CONJUGATION

通过谷胱甘肽结合激活促癌剂

• 批准号: 6174230
• 负责人: JAMES B WHEELER
• 金额: $ 3.75万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-03 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6174230
• 关键词: DNA damage; Salmonella typhimurium; adduct; carcinogens; chemical conjugate; chemical synthesis; enzyme mechanism; glutathione transferase; haloalkane; mutagen testing; mutagens; oligonucleotides; protein purification

Industrial compounds such as methylene chloride, dibromoethane, and butadiene are known carcinogens in rodents. These compounds are activated by conjugation with glutathione (GSH) and cause mutations in DNA by forming DNA adducts. The intermediates of this reaction are believed to be glutathione conjugates formed by glutathione S- transferases (GSTs). This proposal compares the ability of rat, human, and bacterial GSTs to form mutagenic DNA adducts with GSH conjugates in the presence of different haloalkanes and butadiene diepoxide. The proposed research will examine the genotoxicity of these compounds, the mechanism of activation by GST, the stability of the compounds, and the DNA adducts formed. The Ames test will be used to determine the mutagenicity of the various compounds when GST is being expressed in Salmonella typhuurium. Recombinant GSTs from human, rat and bacterial sources will be purified and used for in vitro assays to determine the mechanism of DNA adduct formation. Chemically synthesized intermediates of the GSH activation reaction will be tested for stability under various conditions and oligonucleotides will be used to test the formation of DNA adducts.

工业化合物，例如氯化甲基，二溴甲烷和 丁二烯是啮齿动物中已知的致癌物。 这些化合物是 通过与谷胱甘肽（GSH）结合激活并引起突变 DNA通过形成DNA加合物。 该反应的中间体是 被认为是谷胱甘肽s-形成的谷胱甘肽结合物 转移酶（GST）。 该建议比较了大鼠人类，人类的能力 和细菌GSTs与GSH共轭形成诱变DNA加合物 存在不同的卤代烷和丁二烯二氧化物的存在。 这 拟议的研究将检查这些化合物的遗传毒性，即 GST激活的机理，化合物的稳定性和 DNA加合物形成。 AMES测试将用于确定 在GST中表达各种化合物的诱变性 沙门氏菌鼠伤寒。 来自人，大鼠和细菌的重组GST 来源将被纯化并用于体外测定法以确定 DNA加合物形成的机理。 化学合成的中间体 GSH激活反应的稳定性将在 各种条件和寡核苷酸将用于测试 DNA加合物的形成。