ABSTRACT The formation of protein carbonyls in the metal-catalyzed oxidation of human serum albumin (HSA) is characterized using a new analytical approach that involves tagging the modification site with multiple hydrazide reagents. Protein carbonyl formation at lysine and arginine residues was catalyzed with copper and iron ions, and the resulting oxidation patterns in HSA are contrasted. A total of 18 modification sites were identified with iron-ion catalysis and 14 with copper-ion catalysis. However, with the more stringent requirement of identification with at least two tagging reagents, the number of validated modification sites drops to 10 for iron and nine for copper. Of the 14 total validated sites, there were only five in common for the two metal ions. The results illustrate the value of using multiple tagging agents and highlight the selective and specific nature of metal-catalyzed protein oxidations.

中文翻译：

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鉴定特定蛋白质羰基化位点的稳健分析方法：人血清白蛋白的金属催化氧化

摘要 在金属催化的人血清白蛋白 (HSA) 氧化过程中，蛋白质羰基化合物的形成使用一种新的分析方法进行表征，该方法涉及用多种酰肼试剂标记修饰位点。赖氨酸和精氨酸残基处的蛋白质羰基形成由铜和铁离子催化，并对比了 HSA 中产生的氧化模式。铁离子催化鉴定了总共18个修饰位点，铜离子催化鉴定了14个。然而，随着对至少两种标记试剂进行鉴定的更严格要求，已验证的修饰位点数量下降到铁的 10 个和铜的 9 个。在总共 14 个经过验证的位点中，两种金属离子只有 5 个相同。