DETECTING OXIDATIVE MODIFICATION OF BIOMOLECULES IN VIVO

检测体内生物分子的氧化修饰

• 批准号: 6336876
• 负责人: JAY W HEINECKE
• 金额: $ 1.3万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-08-01 至 2001-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6336876
• 关键词: animal tissue; bioengineering /biomedical engineering; biological products; biomedical resource; spectrometry; technology /technique

Oxidative modification of low density lipoprotein (LDL) is thought to participate in the pathogenesis of atherosclerosis, but the pathways involved in such modification in vivo are poorly understood. The enzyme myeloperoxidase (MPO) is present in catalytically active form in human atherosclerotic lesions, and MPO can use a variety of substrates, including chloride, tyrosine, and nitrite, to generate oxidants that react with molecules in their environment. We have used mass spectrometry to identify and quantitate products of such reactions, including chlorinated cholesterol species, 3-chlorotyrosine, o-dityrosine, 3-nitrotyrosine, a family of reactive aldehydes derived from a-amino acids, and a lysine derivative formed by reaction with such aldehydes. We have developed isotope dilution GC/MS assays for such products that exploit the great sensitivity of negative ion electron capture MS analyses. These assays have permitted us to measure these products in cellular sys tems in vit ro and in LDL isolated from plasma and from human

低密度脂蛋白（LDL）的氧化修饰被认为 参与动脉粥样硬化的发病机制，但 人们对体内这种修饰所涉及的途径知之甚少。 髓过氧化物酶 (MPO) 存在于催化活性物质中 人类动脉粥样硬化病变中形成，MPO可利用多种 底物，包括氯化物、酪氨酸和亚硝酸盐，以产生 与环境中的分子发生反应的氧化剂。 我们已经用过 质谱法来识别和定量此类产品 反应，包括氯化胆固醇种类， 3-氯酪氨酸、邻二酪氨酸、3-硝基酪氨酸，一类反应性酪氨酸 由α-氨基酸衍生的醛，以及形成的赖氨酸衍生物 通过与此类醛反应。 我们开发了同位素稀释 利用 GC/MS 的高灵敏度对此类产品进行分析 负离子电子捕获质谱分析。 这些测定有 允许我们在体外的细胞系统中测量这些产品 以及从血浆和人体中分离出的低密度脂蛋白