酪氨酸
硝化作用
氧化应激
生物化学
硝基酪氨酸
生物
氧化磷酸化
磷酸蛋白质组学
蛋白质酪氨酸磷酸酶
化学
酶
蛋白质磷酸化
蛋白激酶A
有机化学
一氧化氮合酶
作者
Jason Porter,Hyo Sang Jang,Elise M. Van Fossen,Duy Nguyen,Taylor Willi,Richard B. Cooley,Ryan A. Mehl
标识
DOI:10.1021/acschembio.9b00371
摘要
Tyrosine nitration has served as a major biomarker for oxidative stress and is present in high abundance in over 50 disease pathologies in humans. While data mounts on specific disease pathways from specific sites of tyrosine nitration, the role of these modifications is still largely unclear. Strategies for installing site-specific tyrosine nitration in target proteins in eukaryotic cells, through routes not dependent on oxidative stress, would provide a powerful method to address the consequences of tyrosine nitration. Developed here is a Methanosarcina barkeri aminoacyl-tRNA synthetase/tRNA pair that efficiently incorporates nitrotyrosine site-specifically into proteins in mammalian cells. We demonstrate the utility of this approach to produce nitrated proteins identified in disease conditions by producing site-specific nitroTyr-containing manganese superoxide dismutase and 14-3-3 proteins in eukaryotic cells.
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