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 从动脉粥样硬化到衰老的各种过程。 事实证明，识别潜在的氧化途径很困难， 然而，由于缺乏不同氧化的特异性标记 途径。 先前的体外研究表明 3-氯酪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充当 体内吞噬细胞介导的组织损伤的指数。 在这里我们 描述一种高度灵敏且特异的分析方法 组织中 3-氯酪氨酸含量的定量。 化验 将气相色谱法与稳定同位素稀释质量相结合 光谱法，它可以检测 3-氯酪氨酸的阿托摩尔水平 单一测定。 此外，该方法具有高度重现性， 样本间和样本内方差系数<3%。 这 3-氯酪氨酸 e 的特异性、敏感性和重现性 确定应该使该方法对于探索角色有用 的 髓过氧化物酶催化体内氧化反应。