Acrolein inhibits cytokine gene expression by alkylating cysteine and arginine residues in the NF-κB1 DNA binding domain
Acrolein inhibits cytokine gene expression by alkylating cysteine and arginine residues in the NF-κB1 DNA binding domain
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DOI：
10.1074/jbc.m611527200
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发表时间：
2007-07-06
影响因子：
4.8
通讯作者：
Freed, Brian M.
中科院分区：
文献类型：
--
作者：
Lambert, Cherie;Li, Jimei;Freed, Brian M.
香烟烟雾是肺部 T 细胞反应的有效抑制剂，导致免疫监视功能下降和呼吸道感染发生率增加。香烟烟雾中的α、β-不饱和醛（丙烯醛和巴豆醛）抑制人类T细胞产生白细胞介素-2（IL-2）、IL-10、粒细胞巨噬细胞集落刺激因子、干扰素-γ和肿瘤坏死因子-α，但不抑制IL-8的产生。香烟烟雾中的饱和醛（乙醛、丙醛和丁醛）不活跃。丙烯醛抑制 T 细胞促有丝分裂刺激后 NF-κ B DNA 结合活性的诱导，但对 NFAT 或 AP-1 的诱导没有影响。在染色质免疫沉淀测定中，丙烯醛抑制 NF-kappa B1 (p50) 与 IL-2 启动子的结合超过 99%。在电泳迁移率变动测定中使用纯化的重组 p50，我们证明丙烯醛在阻断 DNA 与 NF-kappa B 共有序列结合方面的效力比巴豆醛强 2000 倍。基质辅助激光解吸/电离飞行时间和串联质谱分析表明，丙烯醛烷基化了 DNA 结合域中的两个氨基酸（Cys-61 和 Arg-307）。巴豆醛与 Cys-61 反应，但不与 Arg-307 反应，而香烟烟雾中的饱和醛不与 p50 反应。这些实验表明，香烟烟雾中的醛可以通过直接修饰转录因子来调节基因表达。
Cigarette smoke is a potent inhibitor of pulmonary T cell responses, resulting in decreased immune surveillance and an increased incidence of respiratory tract infections. The alpha, beta-unsaturated aldehydes in cigarette smoke ( acrolein and crotonaldehyde) inhibited production of interleukin- 2 (IL-2), IL-10, granulocyte-macrophage colony-stimulating factor, interferon-gamma, and tumor necrosis factor-alpha by human T cells but did not inhibit production of IL-8. The saturated aldehydes (acetaldehyde, propionaldehyde, and butyraldehyde) in cigarette smoke were inactive. Acrolein inhibited induction of NF-kappa B DNA binding activity after mitogenic stimulation of T cells but had no effect on induction of NFAT or AP-1. Acrolein inhibited NF-kappa B1 (p50) binding to the IL-2 promoter in a chromatin immunoprecipitation assay by > 99%. Using purified recombinant p50 in an electrophoretic mobility shift assay, we demonstrated that acrolein was 2000-fold more potent than crotonaldehyde in blocking DNA binding to an NF-kappa B consensus sequence. Matrix-assisted laser desorption/ionization time-of-flight and tandem mass spectrometry demonstrated that acrolein alkylated two amino acids (Cys-61 and Arg-307) in the DNA binding domain. Crotonaldehyde reacted with Cys-61, but not Arg-307, whereas the saturated aldehydes in cigarette smoke did not react with p50. These experiments demonstrate that aldehydes in cigarette smoke can regulate gene expression by direct modification of a transcription factor.