Quantitative analysis of redox-sensitive proteome with DIGE and ICAT

Cexiong Fu, Jun Hu, Tong Liu, Tetsuro Ago, Junichi Sadoshima, Hong Li

Research output: Contribution to journalArticle

80 Citations (Scopus)

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-κ:B-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.

Original languageEnglish
Pages (from-to)3789-3802
Number of pages14
JournalJournal of Proteome Research
Volume7
Issue number9
DOIs
Publication statusPublished - Sep 1 2008

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Proteome
Electrophoresis
Isotopes
Oxidation-Reduction
Gels
Chemical analysis
Proteins
Sulfhydryl Compounds
Voltage-Dependent Anion Channel 2
Voltage-Dependent Anion Channels
Epoxide Hydrolases
Oxidation
Reticulum
Calcium-Transporting ATPases
Biological systems
Proteomics
Cysteine
Membrane Proteins
Tissue

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Chemistry(all)

Cite this

Quantitative analysis of redox-sensitive proteome with DIGE and ICAT. / Fu, Cexiong; Hu, Jun; Liu, Tong; Ago, Tetsuro; Sadoshima, Junichi; Li, Hong.

In: Journal of Proteome Research, Vol. 7, No. 9, 01.09.2008, p. 3789-3802.

Research output: Contribution to journalArticle

Fu, Cexiong ; Hu, Jun ; Liu, Tong ; Ago, Tetsuro ; Sadoshima, Junichi ; Li, Hong. / Quantitative analysis of redox-sensitive proteome with DIGE and ICAT. In: Journal of Proteome Research. 2008 ; Vol. 7, No. 9. pp. 3789-3802.
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