Site-specific identification and validation of hepatic histone nitration in vivo: Implications for alcohol-induced liver injury.
Site-specific identification and validation of hepatic histone nitration in vivo: Implications for alcohol-induced liver injury.
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体内肝组蛋白硝化的位点特异性鉴定和验证：对酒精性肝损伤的影响。
DOI：
10.1002/jms.4713
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发表时间：
2021
期刊：
影响因子：
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通讯作者：
StevensJr,StanleyM
中科院分区：
文献类型：
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作者：
Kriss,CrystinaL;Duro,Nalvi;Nadeau,OwenW;Guergues,Jennifer;Chavez-Chiang,Omar;Culver-Cochran,AshleyE;Chaput,Dale;Varma,Sameer;StevensJr,StanleyM
氧化应激和硝化应激涉及多种生物过程和疾病状态的分子机制，包括癌症、衰老、心血管疾病、神经系统疾病、糖尿病和酒精诱导的肝损伤。硝化应激的一个标志是3 -硝基酪氨酸或蛋白酪氨酸硝化（PTN）的形成，这在炎症和组织损伤期间已被观察到；然而，PTN在疾病进展中的作用或可能的发病机制仍不清楚。我们在酒精诱导的肝损伤模型中发现，慢性酒精暴露（28天）后，野生型雄性C57BL/6J小鼠肝脏内肝细胞核中PTN增加。分离肝核的高分辨率质谱分析揭示了组蛋白上酪氨酸硝化的几个新位点。组蛋白硝化位点通过串联质谱（MS/MS）分析与体内鉴定的各自硝基酪氨酸位点序列等效的代表性合成硝化肽进行验证。我们进一步研究了使用分子动力学（MD）模拟确定的新型组蛋白H3 Tyr41 （H3Y41）硝化位点的潜在结构影响。组蛋白H3Y41硝化和非硝化形式的MD模拟显示，H3Y41硝化后，DNA界面的结构发生了显著变化。本研究的结果表明，除了组蛋白上发生的其他已知的翻译后修饰（例如乙酰化和甲基化）外，PTN还可以诱导染色质结构变化，可能影响与酒精诱导肝损伤发展相关的基因转录过程。
Oxidative and nitrative stress have been implicated in the molecular mechanisms underlying a variety of biological processes and disease states including cancer, aging, cardiovascular disease, neurological disorders, diabetes, and alcohol‐induced liver injury. One marker of nitrative stress is the formation of 3‐nitrotyrosine, or protein tyrosine nitration (PTN), which has been observed during inflammation and tissue injury; however, the role of PTN in the progression or possibly the pathogenesis of disease is still unclear. We show in a model of alcohol‐induced liver injury that an increase in PTN occurs in hepatocyte nuclei within the liver of wild‐type male C57BL/6J mice following chronic ethanol exposure (28 days). High‐resolution mass spectrometric analysis of isolated hepatic nuclei revealed several novel sites of tyrosine nitration on histone proteins. Histone nitration sites were validated by tandem mass spectrometry (MS/MS) analysis of representative synthetic nitropeptides equivalent in sequence to the respective nitrotyrosine sites identified in vivo. We further investigated the potential structural impact of the novel histone H3 Tyr41 (H3Y41) nitration site identified using molecular dynamics (MD) simulations. MD simulations of the nitrated and non‐nitrated forms of histone H3Y41 showed significant structural changes at the DNA interface upon H3Y41 nitration. The results from this study suggest that, in addition to other known post‐translational modifications that occur on histone proteins (e.g., acetylation and methylation), PTN could induce chromatin structural changes, possibly affecting gene transcription processes associated with the development of alcohol‐induced liver injury.