Proteomic Characterization of the Cellular Response to Nitrosative Stress Mediated by S-nitrosoglutathione Reductase Inhibition
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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 "nitrosative stress". 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)-including heme oxygenase 1 (HO-1) and glutamate cysteine ligase modulatory subunit-were 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.
Hayashida K, Bagchi A, Miyazaki Y, Hirai S, Seth D, Silverman M Circulation. 2018; 139(6):815-827.
PMID: 30586713 PMC: 6361681. DOI: 10.1161/CIRCULATIONAHA.117.032488.
Cox N, Luo P, Smith T, Bisnett B, Soderblom E, Boyce M Front Endocrinol (Lausanne). 2018; 9:606.
PMID: 30459710 PMC: 6232944. DOI: 10.3389/fendo.2018.00606.
Yang Z, Bochkov Y, Voelker D, Foster M, Que L Am J Respir Cell Mol Biol. 2018; 60(1):58-67.
PMID: 30156431 PMC: 6348719. DOI: 10.1165/rcmb.2018-0058OC.
Proteome profiling of s-nitrosylated synaptosomal proteins by isobaric mass tags.
Wijasa T, Sylvester M, Brocke-Ahmadinejad N, Kummer M, Brosseron F, Gieselmann V J Neurosci Methods. 2017; 291:95-100.
PMID: 28789995 PMC: 5625850. DOI: 10.1016/j.jneumeth.2017.08.005.
In-depth proteomic profiling of left ventricular tissues in human end-stage dilated cardiomyopathy.
Liu S, Xia Y, Liu X, Wang Y, Chen Z, Xie J Oncotarget. 2017; 8(29):48321-48332.
PMID: 28427148 PMC: 5564650. DOI: 10.18632/oncotarget.15689.