Nitration and nitrosation of N-acetyl-L-tryptophan and tryptophan residues in proteins by various reactive nitrogen species.
Nitration and nitrosation of N-acetyl-L-tryptophan and tryptophan residues in proteins by various reactive nitrogen species.
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DOI：
10.1016/j.freeradbiomed.2004.05.030
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发表时间：
2004-09
影响因子：
7.4
通讯作者：
Toshinori Suzuki;H. Mower;M. Friesen;I. Gilibert;T. Sawa;H. Ohshima
中科院分区：
文献类型：
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作者：
Toshinori Suzuki;H. Mower;M. Friesen;I. Gilibert;T. Sawa;H. Ohshima
蛋白质是活性氮物质（例如过氧亚硝酸盐和二氧化氮）的目标。在蛋白质的各种氨基酸中，色氨酸残基特别容易受到活性氮的攻击。我们在不同条件下进行了过氧亚硝酸盐和其他活性氮与N-乙酰基-L-色氨酸的反应实验。根据质谱和紫外光谱，四种主要产物被鉴定为1-亚硝基-N-乙酰基-L-色氨酸、1-硝基-N-乙酰基-L-色氨酸、6-硝基-N-乙酰基-L-色氨酸和N-乙酰基-N'-甲酰基-L-犬尿氨酸。与 SIN-1（过氧亚硝酸盐生成剂）、Angeli 盐（硝酰基供体）和精胺 NONOate（一氧化氮供体）的反应生成亚硝基衍生物，但不生成硝基衍生物。髓过氧化物酶-H2O2-NO2-系统产生硝基衍生物，但不产生亚硝基衍生物。在生理条件下，6-硝基-N-乙酰基-L-色氨酸是稳定的，而1-亚硝基和1-硝基衍生物则分解，半衰期分别为1.5和18小时。用各种活性氮处理后，对牛血清白蛋白进行酶水解，并通过 HPLC 和电化学检测分析 6-硝基-L-色氨酸和 3-硝基-L-酪氨酸。硝化蛋白质中 6-硝基-L-色氨酸和 3-硝基-L-酪氨酸的水平相似。蛋白质中的 6-硝基-L-色氨酸可以作为蛋白质硝化的附加生物标志物进行测量。
Proteins are targets of reactive nitrogen species such as peroxynitrite and nitrogen dioxide. Among the various amino acids in proteins, tryptophan residues are especially susceptible to attack by reactive nitrogen species. We carried out experiments on the reactions of peroxynitrite and other reactive nitrogen species with N-acetyl-l-tryptophan under various conditions. Four major products were identified as 1-nitroso-N-acetyl-l-tryptophan, 1-nitro-N-acetyl-l-tryptophan, 6-nitro-N-acetyl-l-tryptophan, and N-acetyl-N′-formyl-l-kynurenine on the basis of their mass and UV spectra. The reactions with SIN-1 (a peroxynitrite generator), Angeli's salt (a nitroxyl donor), and spermine NONOate (a nitric oxide donor) generated the nitroso derivative but not the nitro derivatives. A myeloperoxidase–H2O2–NO2−system generated the nitro derivatives but not the nitroso derivative. Under physiological conditions 6-nitro-N-acetyl-l-tryptophan was stable, whereas the 1-nitroso and 1-nitro derivatives decomposed with half-lives of 1.5 and 18 h, respectively. After treatment with various reactive nitrogen species, bovine serum albumin was enzymatically hydrolyzed and analyzed for 6-nitro-l-tryptophan and 3-nitro-l-tyrosine by HPLC with electrochemical detection. Levels of 6-nitro-l-tryptophan and 3-nitro-l-tyrosine were similar in the nitrated protein. 6-Nitro-l-tryptophan in proteins can be measured as an additional biomarker of protein nitration.