Abstract
The advent of sensitive and robust quantitative proteomics techniques has been emerging as a vital tool for deciphering complex biological puzzles that would have been challenging to conventional molecular biology methods. The method here describes the use of two isotope labeling techniques—isobaric tags for relative and absolute quantification (iTRAQ) and redox isotope-coded affinity tags (ICAT)—to elucidate the cardiovascular redox-proteome changes and thioredoxin 1 (Trx1)-regulated protein network in cardiac-specific Trx1 transgenic mouse models. The strategy involves the use of an amine-labeling iTRAQ technique, gauging the global proteome changes in Trx1 transgenic mice at the protein level, while ICAT, labeling redox-sensitive cysteines, reveals the redox status of cysteine residues. Collectively, these two quantitative proteomics techniques can not only quantify global changes of the cardiovascular proteome but also pinpoint specific redox-sensitive cysteine sites that are subjected to Trx1-catalyzed reduction.
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Acknowledgements
The authors are grateful for the funding support from NIH/NINDS grant P30 NS046593 to H.L for the support of a NINDS NeuroProteomics Core Facility at UMDNJ-New Jersey Medical School.
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Fu, C., Liu, T., Parrott, A.M., Li, H. (2013). Identification of Thioredoxin Target Protein Networks in Cardiac Tissues of a Transgenic Mouse. In: Vivanco, F. (eds) Heart Proteomics. Methods in Molecular Biology, vol 1005. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-386-2_15
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DOI: https://doi.org/10.1007/978-1-62703-386-2_15
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Publisher Name: Humana Press, Totowa, NJ
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