MASS SPECTROMETRIC QUANTIFICATION OF 3 CHLOROTYROSINE IN HUMAN TISSUES

人体组织中 3 氯酪氨酸的质谱定量

• 批准号: 6486664
• 负责人: STANLEY HAZEN
• 金额: $ 15.75万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6486664
• 关键词: biomedical resource; cardiovascular system; enzymes; gas; human tissue; minerals; proteins; spectrometry; technology /technique

Oxidative modification of proteins has been implicated in a variety of processes ranging from atherosclerosis to aging. Identifying the underlying oxidation pathways has proven difficult, however, due to the lack of specific markers for distinct oxidation pathways. Previous in vitro studies demonstrated that 3-chlorotyrosine is a specific product of myeloperoxidase-catalyzed oxidative damage and that the chlorinated amino acid may thus serve as an index of phagocyte-mediated tissue injury in vivo. Here we describe a highly sensitive and specific analytical method for the quantification of 3-chlorotyrosine content of tissues. The assay combines gas chromatography with stable isotope dilution mass spectrometry, and it detects attomole levels of 3-chlrotyrosine in a single determination. Furthermore, the method is highly reproducible, with inter- and intra-sample coefficients of variance of <3%. The specificity, sensitivity, and reproducibility of 3-chlorotyrosin e determin ation should make this method useful for exploring the role of myeloperoxidase in catalyzing oxidative reactions in vivo.

蛋白质的氧化修饰与A有关 从动脉粥样硬化到衰老的各种过程。 识别潜在的氧化途径已被证明很困难， 但是，由于缺乏明显氧化的特定标记 途径。 以前的体外研究表明 3-氯尼罗氨酸是髓过氧化物酶催化的特定产物 氧化损伤，因此氯化氨基酸可以作为 吞噬细胞介导的组织损伤的指数。 我们在这里 描述一种高度敏感和特定的分析方法 定量组织的3-氯尼罗尼含量。 测定法 结合气相色谱与稳定的同位素稀释量 光谱法检测到A 单个确定。 此外，该方法是高度可重复的， 差异和样本内的方差<3％。 这 3-氯曲霉素E的特异性，灵敏度和可重复性 确定应使此方法对于探索角色有用 的 在体内催化氧化反应中的骨髓过氧化物酶。