Publication Information
Liu et al., 2014
No external accession available
Abstract
Proteomics. 2014 Mar;14(6):750-62. doi: 10.1002/pmic.201300307. Epub 2014 Jan
28.
Identification of redox-sensitive cysteines in the Arabidopsis proteome using
OxiTRAQ, a quantitative redox proteomics method.
Liu P(1), Zhang H, Wang H, Xia Y.
Author information:
(1)Department of Biology, Hong Kong Baptist University, Hong Kong, P. R. China.
Cellular 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.
DOI: 10.1002/pmic.201300307
PMID: 24376095 [Indexed for MEDLINE]
