Uppal, Karan Soltow, Quinlyn A Strobel, Frederick H et al. (2013) High-performance metabolic profiling with dual chromatography-Fourier-transform mass spectrometry (DC-FTMS) for study of the exposome. Soltow, Quinlyn A Strobel, Frederick H Mansfield, Keith G et al. Nuclear and Cytoplasmic Redox in Oxidative Stress Nuclear and cytoplasmic redox in oxidative stress Successful completion of these aims will quantitatively define the redox of nuclear and cytoplasmic Trx, show whether Trx and GSH have distinct functions in cell signaling and oxidative stress, and test whether nuclear or cytoplasmic Trx redox is critical in transcriptional activation during oxidant signaling. This will be done with retroviral constructs containing mitochondrial Trx reductase in which the mitochondrial targeting sequence has been removed and a nuclear import or export signal has been added. The third aim i s to selectively modify Trx redox in nuclear and cytoplasmic compartments to examine cause-effect relationships between Trx redox and transcriptional activation. The second aim i s to determine whether nuclear translocation of Nrf2, a transcription factor activated by oxidative stress and implicated in regulation of detoxification gene expression, is specifically associated with oxidation of cytoplasmic Trx redox while nuclear Trx remains reduced. The first aim i s to determine the redox state of nuclear and cytoplasmic Trx in HT29 cells during cell growth, differentiation and apoptosis. Redox of cellular GSH/GSSG and NADPH/NADP+ will be simultaneously determined to gain insight into the control and interaction of these redox pools. The purpose of this proposal is to use a novel Redox-Western blot technique to investigate the redox of Trx in cells, and use this in combination with cell fractionation to measure redox of the nuclear and cytoplasmic Trx pools. Although a considerable amount of information exists on the redox responses of the GSH system during redox signaling and oxidative stress, little information is available on the redox of Trx. However, there are also important differences, such as the role of thioredoxin in maintaining transcription factors in their reduced and functional forms. These systems have similar NADPH-dependent reductases and have overlapping functions and activities. Two major thiol-containing defense systems exist to protect against oxidative stress, one utilizing the tripeptide glutathione (GSH) and the other using a small protein, thioredoxin (Trx). Oxidative stress is an important component of toxicity due to environmental, occupational and therapeutic agents.
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