Identification of redox-sensitive cysteines in the arabidopsis proteome using OxiTRAQ, a quantitative redox proteomics method
KAUST DepartmentBioscience Core Lab
Online Publication Date2014-01-28
Print Publication Date2014-03
Permanent link to this recordhttp://hdl.handle.net/10754/563356
MetadataShow full item record
AbstractCellular redox status plays a key role in mediating various physiological and developmental processes often through modulating activities of redox-sensitive proteins. Various stresses trigger over-production of reactive oxygen/nitrogen species which lead to oxidative modifications of redox-sensitive proteins. Identification and characterization of redox-sensitive proteins are important steps toward understanding molecular mechanisms of stress responses. Here, we report a high-throughput quantitative proteomic approach termed OxiTRAQ for identifying proteins whose thiols undergo reversible oxidative modifications in Arabidopsis cells subjected to oxidative stress. In this approach, a biotinylated thiol-reactive reagent is used for differential labeling of reduced and oxidized thiols. The biotin-tagged peptides are affinity purified, labeled with iTRAQ reagents, and analyzed using a paralleled HCD-CID fragmentation mode in an LTQ-Orbitrap. With this approach, we identified 195 cysteine-containing peptides from 179 proteins whose thiols underwent oxidative modifications in Arabidopsis cells following the treatment with hydrogen peroxide. A majority of those redox-sensitive proteins, including several transcription factors, were not identified by previous redox proteomics studies. This approach allows identification of the specific redox-regulated cysteine residues, and offers an effective tool for elucidation of redox proteomes. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
SponsorsThis work was supported by Research Grants Council of Hong Kong (grant nos. HKBU1/CRF/10 and HKBU261910 to Y.X.) and by HKBU Strategic Development Fund (to Y.X.).
- Proteomic analysis of early-responsive redox-sensitive proteins in Arabidopsis.
- Authors: Wang H, Wang S, Lu Y, Alvarez S, Hicks LM, Ge X, Xia Y
- Issue date: 2012 Jan 1
- Quantifying reversible oxidation of protein thiols in photosynthetic organisms.
- Authors: Slade WO, Werth EG, McConnell EW, Alvarez S, Hicks LM
- Issue date: 2015 Apr
- Isotope-coded affinity tag (ICAT) approach to redox proteomics: identification and quantitation of oxidant-sensitive cysteine thiols in complex protein mixtures.
- Authors: Sethuraman M, McComb ME, Huang H, Huang S, Heibeck T, Costello CE, Cohen RA
- Issue date: 2004 Nov-Dec
- Quantitative redox proteomics: the NOxICAT method.
- Authors: Lindemann C, Leichert LI
- Issue date: 2012
- Differential redox proteomics allows identification of proteins reversibly oxidized at cysteine residues in endothelial cells in response to acute hypoxia.
- Authors: Izquierdo-Álvarez A, Ramos E, Villanueva J, Hernansanz-Agustín P, Fernández-Rodríguez R, Tello D, Carrascal M, Martínez-Ruiz A
- Issue date: 2012 Sep 18