Identification of redox-sensitive cysteines in the arabidopsis proteome using OxiTRAQ, a quantitative redox proteomics method

Handle URI:
http://hdl.handle.net/10754/563356
Title:
Identification of redox-sensitive cysteines in the arabidopsis proteome using OxiTRAQ, a quantitative redox proteomics method
Authors:
Liu, Pei; Zhang, Huoming ( 0000-0001-5416-0358 ) ; Wang, Hai; Xia, Yiji
Abstract:
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.
KAUST Department:
Biosciences Core Lab
Publisher:
Wiley-Blackwell
Journal:
PROTEOMICS
Issue Date:
28-Jan-2014
DOI:
10.1002/pmic.201300307
PubMed ID:
24376095
Type:
Article
ISSN:
16159853
Sponsors:
This 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.).
Appears in Collections:
Articles; Bioscience Core Lab

Full metadata record

DC FieldValue Language
dc.contributor.authorLiu, Peien
dc.contributor.authorZhang, Huomingen
dc.contributor.authorWang, Haien
dc.contributor.authorXia, Yijien
dc.date.accessioned2015-08-03T11:46:34Zen
dc.date.available2015-08-03T11:46:34Zen
dc.date.issued2014-01-28en
dc.identifier.issn16159853en
dc.identifier.pmid24376095en
dc.identifier.doi10.1002/pmic.201300307en
dc.identifier.urihttp://hdl.handle.net/10754/563356en
dc.description.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.en
dc.description.sponsorshipThis 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.).en
dc.publisherWiley-Blackwellen
dc.subjectArabidopsis thalianaen
dc.subjectITRAQen
dc.subjectOxidative stressen
dc.subjectOxiTRAQen
dc.subjectPlant proteomicsen
dc.subjectRedox-sensitive proteinen
dc.subjectRedoxomeen
dc.titleIdentification of redox-sensitive cysteines in the arabidopsis proteome using OxiTRAQ, a quantitative redox proteomics methoden
dc.typeArticleen
dc.contributor.departmentBiosciences Core Laben
dc.identifier.journalPROTEOMICSen
dc.contributor.institutionDepartment of Biology, Hong Kong Baptist University, Hong Kong, Chinaen
dc.contributor.institutionPartner State Key Laboratory of Agrobiotechnology, The Chinese University of Hong Kong, Hong Kong, Chinaen
kaust.authorZhang, Huomingen

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