INTERACTION OF CHROMATE WITH MITOCHONDRIA

铬酸盐与线粒体的相互作用

• 批准号: 3172679
• 负责人: KAREN E WETTERHAHN
• 金额: $ 8.45万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-06-01 至 1986-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3172679
• 关键词: chromium; cytochrome P450; cytotoxicity; density gradient ultracentrifugation; electron spin resonance spectroscopy; gel electrophoresis; metal metabolism; metal oxide; mitochondria; nucleic acid denaturation; nucleic acid structure; radiotracer

The objective of this research project is to study the chromate-reductase activity of mitochondria, to determine the damage induced in mitochondrial DNA by chromate and to determine the effect of chromium on DNA structure. The mitochondria will serve as a model system for studying the uptake-reduction model for chromate carcinogenicity. The components of mitochondria, such as the electron transport chain or the cytochrome P-450-like activity, which are responsible for chromate metabolism will be determined using inhibitors and purified components. The generation of reactive species such as free radicals or chromium (V) during mitochondrial chromate reduction will be monitored by EPR spectroscopy. The ability of chromium to affect supercoiling and bind to specific sequences of mitochondrial (mt) DNA and pBR322 DNA will be studied by buoyant density gradient, gel electrophoresis, restriction enduclease digestion and DNA sequencing techniques. The ability of chromium to affect the B-Z DNA transition of poly (dG-dC) . poly (dG-dC) will be determined by circular dichroism. The ability of chromium to condense DNA into PsiDNA or toroidal structures will be determined by circular dichroism and sedimentation studies. Local denaturation of DNA by bound chromium will be examined by S1 nuclease sensitivity assay.

本研究项目的目的是研究铬酸盐还原酶。 线粒体的活性，以确定线粒体的损伤 用铬酸盐处理DNA，并测定铬对DNA结构的影响。 线粒体将作为一个模型系统来研究 铬酸盐致癌的摄取-还原模型。的组件 线粒体，如电子传输链或细胞色素 负责铬酸盐代谢的P-450样活性将是 使用抑制剂和纯化组分进行测定。新一代人 线粒体中的活性物种，如自由基或铬(V) 铬酸盐还原将通过EPR光谱进行监测。的能力 影响超级卷曲并与特定的序列结合的铬 线粒体(Mt)DNA和pBR322 DNA将通过浮力密度进行研究 梯度、凝胶电泳、限制性内切酶消化和DNA 测序技术。铬对B-Z DNA的影响 聚(DG-DC)的转变。聚(DG-DC)将通过循环测定 二向色性。铬使DNA凝聚成PsiDNA或环状DNA的能力 构造将通过圆二向色性和沉积作用来确定 学习。结合铬对DNA的局部变性将通过 S1核酸酶敏感试验。