title: Quantitative analysis of redox-sensitive proteome with DIGE and ICAT.
abstract: Oxidative modifications of protein thiols are important mechanisms for regulating protein functions. The present study aimed to compare the relative effectiveness of two thiol-specific quantitative proteomic techniques, difference gel electrophoresis (DIGE) and isotope coded affinity tag (ICAT), for the discovery of redox-sensitive proteins in heart tissues. We found that these two methods were largely complementary; each could be used to reveal a set of unique redox-sensitive proteins. Some of these proteins are low-abundant signaling proteins and membrane proteins. From DIGE analysis, we found that both NF-kappaB-repressing protein and epoxide hydrolase were sensitive to H 2O 2 oxidation. In ICAT analysis, we found that specific cysteines within sacroplasmic endoplamic reticulum calcium ATPase 2 and voltage-dependent anion-selective channel protein 1 were sensitive to H 2O 2 oxidation. From these analyses, we conclude that both methods should be employed for proteome-wide studies, to maximize the possibility of identifying proteins containing redox-sensitive cysteinyl thiols in complex biological systems.
authors.affiliation: Center for Advanced Proteomics Research and Department of Biochemistry and Molecular Biology, UMDNJ-New Jersey Medical School Cancer Center, Newark, New Jersey 07103, USA.
authors.forename: Cexiong
authors.initials: C
authors.lastname: Fu
authors.forename: Jun
authors.initials: J
authors.lastname: Hu
authors.forename: Tong
authors.initials: T
authors.lastname: Liu
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Ago
authors.forename: Junichi
authors.initials: J
authors.lastname: Sadoshima
authors.forename: Hong
authors.initials: H
authors.lastname: Li
mesh_terms: D000345:Affinity Labels; D000595:Amino Acid Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D006861:Hydrogen Peroxide / Q000737:chemistry; D009124:Muscle Proteins / Q000737:chemistry; D009206:Myocardium / Q000737:chemistry; D010084:Oxidation-Reduction; D020543:Proteome
chemical_list: D000345:Affinity Labels; D009124:Muscle Proteins; D020543:Proteome; D006861:Hydrogen Peroxide