title: Identification of thioredoxin target protein networks in cardiac tissues of a transgenic mouse.
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.
authors.affiliation: Pharmacokinetics, Dynamics and Metabolism, Pfizer Global Research and Development, Pfizer Inc., Groton, CT, USA.
authors.forename: Cexiong
authors.initials: C
authors.lastname: Fu
authors.forename: Tong
authors.initials: T
authors.lastname: Liu
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Parrott
authors.forename: Hong
authors.initials: H
authors.lastname: Li
mesh_terms: D000818:Animals; D056148:Chromatography, Reverse-Phase; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism; D006352:Heart Ventricles / Q000737:chemistry / Q000378:metabolism*; D007553:Isotope Labeling / Q000379:methods*; D051379:Mice; D008822:Mice, Transgenic; D010084:Oxidation-Reduction; D010449:Peptide Mapping; D060066:Protein Interaction Maps; D059748:Proteolysis; D020543:Proteome / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013194:Staining and Labeling / Q000379:methods*; D013879:Thioredoxins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014357:Trypsin / Q000737:chemistry
chemical_list: D020543:Proteome; C515559:Txn1 protein, mouse; D013879:Thioredoxins; D014357:Trypsin; D003545:Cysteine