@inbook{4c116b2d3ad44a02baa23921d5d5b3f8,
title = "Identification of Thioredoxin Target Disulfides Using Isotope-Coded Affinity Tags",
abstract = "Thioredoxins (Trx) are small redox proteins that reduce disulfide bonds in various target proteins and maintain cellular thiol redox control. Here, a thiol-specific labeling and affinity enrichment approach for identification and relative quantification of Trx target disulfides in complex protein extracts is described. The procedure utilizes the isotope-coded affinity tag (ICAT) reagents containing a thiol reactive iodoacetamide group and a biotin affinity tag to target peptides containing reduced cysteine residues. The identification of substrates for Trx and the extent of target disulfide reduction is determined by LC-MS/MS-based quantification of tryptic peptides labeled with {"}light{"} (12C) and {"}heavy{"} (13C) ICAT reagents. The methodology can be adapted to monitor the effect of different reductants or oxidants on the redox status of thiol/disulfide proteomes in biological systems.",
keywords = "Cysteine, Disulfide, Iodoacetamide, Isotope-coded affinity tag, Redox proteomics, Thiol, Thioredoxin",
author = "Per H{\"a}gglund and Jakob Bunkenborg and Kenji Maeda and Christine Finnie and Birte Svensson",
year = "2014",
doi = "10.1007/978-1-62703-631-3_47",
language = "English",
isbn = "978-1-62703-630-6",
volume = "Part VIII",
series = "Methods in Molecular Biology",
publisher = "Humana Press",
pages = "677--685",
booktitle = "Plant Proteomics",
}