Nitric oxide (NO) is a potent mediator produced by many mammalian cells. NO undergoes numerous reactions producing several redox forms. NO can act as a cellular signaling molecule, as a damaging species, or as an antioxidant. This depends on concentration, duration of exposure, and on which targets with which it interacts. Among cellular targets, modification of cellular thiols has been shown to be an important signaling mechanism for NO redox forms. This may lead to S-nitrosoglutathione formation or to protein S-nitrosylation. These modifications of cellular thiols can be considered an """"""""on"""""""" signal. This proposal focuses on reverse NO-mediated thiol modification as a potential """"""""off"""""""" signal. Preliminary experiments have found that NO modification of protein thiols leads to formation of protein mixed disulfides, which are known to be effectively reduced by the combined activities of glutaredoxin and the GSH redox cycle. The applicant proposes that this mechanism is the off signal. As such, the GSH redox cycle may play a protective role through the same mechanism. The goals of the proposal are to investigate mechanisms for interaction between NO, cellular thiols and the GHS redox cycle using endothelial cells exposed to exogenous NO donors or cell models in which NO is produced endogenously. The overall hypothesis is that the GSH redox cycle regulates NO mediated signaling by interacting directly with NO related species and by reversing protein thiol modification mechanisms involving protein disulfide formation. It is assumed that similar mechanisms protect cells from the damaging effects of NO.
| Li, Sheng; Wang, Hua; Xian, Ming et al. (2012) Identification of protein nitrosothiols using phosphine-mediated selective reduction. Nitric Oxide 26:20-6 |
| Granillo, Olivia M; Brahmajothi, Mulugu V; Li, Sheng et al. (2008) Pulmonary alveolar epithelial uptake of S-nitrosothiols is regulated by L-type amino acid transporter. Am J Physiol Lung Cell Mol Physiol 295:L38-43 |
| Zhu, Jun; Li, Sheng; Marshall, Zermeena M et al. (2008) A cystine-cysteine shuttle mediated by xCT facilitates cellular responses to S-nitrosoalbumin. Am J Physiol Cell Physiol 294:C1012-20 |
| Buckley, Barbara J; Li, Sheng; Whorton, A Richard (2008) Keap1 modification and nuclear accumulation in response to S-nitrosocysteine. Free Radic Biol Med 44:692-8 |
| Buckley, Barbara J; Marshall, Zermeena M; Whorton, A R (2003) Nitric oxide stimulates Nrf2 nuclear translocation in vascular endothelium. Biochem Biophys Res Commun 307:973-9 |
| Brar, Sukhdev S; Corbin, Zachary; Kennedy, Thomas P et al. (2003) NOX5 NAD(P)H oxidase regulates growth and apoptosis in DU 145 prostate cancer cells. Am J Physiol Cell Physiol 285:C353-69 |
| Brar, Sukhdev S; Kennedy, Thomas P; Sturrock, Anne B et al. (2002) An NAD(P)H oxidase regulates growth and transcription in melanoma cells. Am J Physiol Cell Physiol 282:C1212-24 |
| Brar, S S; Kennedy, T P; Whorton, A R et al. (2001) Reactive oxygen species from NAD(P)H:quinone oxidoreductase constitutively activate NF-kappaB in malignant melanoma cells. Am J Physiol Cell Physiol 280:C659-76 |
| Buckley, B J; Whorton, A R (2000) Adaptive responses to peroxynitrite: increased glutathione levels and cystine uptake in vascular cells. Am J Physiol Cell Physiol 279:C1168-76 |
