Proteomic Characterization of the Cellular Response to Nitrosative Stress Mediated by S-Nitrosoglutathione Reductase Inhibition

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• 作者：Matthew W. Foster ; Zhonghui Yang ; David M. Gooden ; J. Will Thompson ; Carol H. Ball ; Meredith E. Turner ; Yongyong Hou ; Jingbo Pi ; M. Arthur Moseley ; Loretta G. Que
• 刊名：Journal of Proteome Research
• 出版年：2012
• 出版时间：April 6, 2012
• 年：2012
• 卷：11
• 期：4
• 页码：2480-2491
• 全文大小：446K
• 年卷期：v.11,no.4(April 6, 2012)
• ISSN：1535-3907

文摘

The S-nitrosoglutathione-metabolizing enzyme, GSNO reductase (GSNOR), has emerged as an important regulator of protein S-nitrosylation. GSNOR ablation is protective in models of asthma and heart failure, raising the idea that GSNOR inhibitors might hold therapeutic value. Here, we investigated the effects of a small molecule inhibitor of GSNOR (GSNORi) in mouse RAW 264.7 macrophages. We found that GSNORi increased protein S-nitrosylation in cytokine-stimulated cells, and we utilized stable isotope labeling of amino acids in cell culture (SILAC) to quantify the cellular response to this 鈥渘itrosative stress鈥? The expression of several cytokine-inducible immunomodulators, including osteopontin, cyclooxygenase-2, and nitric oxide synthase isoform 2 (NOS2), were decreased by GSNORi. In addition, selective targets of the redox-regulated transcription factor, nuclear factor (erythroid-derived 2)-like 2 (Nrf2)鈥攊ncluding heme oxygenase 1 (HO-1) and glutamate cysteine ligase modulatory subunit鈥攚ere induced by GSNORi in a NOS2- and Nrf2-dependent manner. In cytokine-stimulated cells, Nrf2 protected from GSNORi-induced glutathione depletion and cytotoxicity and HO-1 activity was required for down-regulation of NOS2. Interestingly, GSNORi also affected a marked increase in NOS2 protein stability. Collectively, these data provide the most complete description of the global effects of GSNOR inhibition and demonstrate several important mechanisms for inducible response to GSNORi-mediated nitrosative stress.

Keywords:

nitric oxide; S-nitrosylation; S-nitrosoglutathione; GSNO reductase; proteomics; heme oxygenase