This Article Full Text Full Text (PDF) All Versions of this Article: 138/4/2233    most recent pp.104.058917v1 Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Related articles in Plant Physiol. Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via ISI Web of Science (27) Citing Articles via Google Scholar Google Scholar Articles by Dixon, D. P. Articles by Edwards, R. Search for Related Content PubMed PubMed Citation Articles by Dixon, D. P. Articles by Edwards, R. Agricola Articles by Dixon, D. P. Articles by Edwards, R. Related Collections Reactive Oxygen Species

BIOCHEMICAL PROCESSES AND MACROMOLECULAR STRUCTURES

Stress-Induced Protein S-Glutathionylation in Arabidopsis¹

School of Biological and Biomedical Sciences, University of Durham, Durham DH1 3LE, United Kingdom

S-Glutathionylation (thiolation) is a ubiquitous redox-sensitive and reversible modification of protein cysteinyl residues that can directly regulate their activity. While well established in animals, little is known about the formation and function of these mixed disulfides in plants. After labeling the intracellular glutathione pool with [³⁵S]cysteine, suspension cultures of Arabidopsis (Arabidopsis thaliana ecotype Columbia) were shown to undergo a large increase in protein thiolation following treatment with the oxidant tert-butylhydroperoxide. To identify proteins undergoing thiolation, a combination of in vivo and in vitro labeling methods utilizing biotinylated, oxidized glutathione (GSSG-biotin) was developed to isolate Arabidopsis proteins/protein complexes that can be reversibly glutathionylated. Following two-dimensional polyacrylamide gel electrophoresis and matrix-assisted laser desorption/ionization time of flight mass spectrometry proteomics, a total of 79 polypeptides were identified, representing a mixture of proteins that underwent direct thiolation as well as proteins complexed with thiolated polypeptides. The mechanism of thiolation of five proteins, dehydroascorbate reductase (AtDHAR1), zeta-class glutathione transferase (AtGSTZ1), nitrilase (AtNit1), alcohol dehydrogenase (AtADH1), and methionine synthase (AtMetS), was studied using the respective purified recombinant proteins. AtDHAR1, AtGSTZ1, and to a lesser degree AtNit1 underwent spontaneous thiolation with GSSG-biotin through modification of active-site cysteines. The thiolation of AtADH1 and AtMetS required the presence of unidentified Arabidopsis proteins, with this activity being inhibited by S-modifying agents. The potential role of thiolation in regulating metabolism in Arabidopsis is discussed and compared with other known redox regulatory systems operating in plants.

² These authors contributed equally to the paper.

³ Present address: Smith and Nephew Research Centre, York Science Park, Heslington, York YO1 5DF, UK.

Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.104.058917.

^* Corresponding author; e-mail robert.edwards{at}durham.ac.uk; fax 0044–191–334–1201.

Received December 23, 2004; returned for revision March 23, 2005; accepted May 3, 2005.

Related articles in Plant Physiol.:

On the Inside

Peter V. Minorsky
Plant Physiol. 2005 138: 1896-1897. [Full Text]  

This article has been cited by other articles:

				L. Michelet, M. Zaffagnini, H. Vanacker, P. Le Marechal, C. Marchand, M. Schroda, S. D. Lemaire, and P. Decottignies In Vivo Targets of S-Thiolation in Chlamydomonas reinhardtii J. Biol. Chem., August 1, 2008; 283(31): 21571 - 21578. [Abstract] [Full Text] [PDF]

				M. Zaffagnini, L. Michelet, V. Massot, P. Trost, and S. D. Lemaire Biochemical Characterization of Glutaredoxins from Chlamydomonas reinhardtii Reveals the Unique Properties of a Chloroplastic CGFS-type Glutaredoxin J. Biol. Chem., April 4, 2008; 283(14): 8868 - 8876. [Abstract] [Full Text] [PDF]

				M. N. Martin, P. H. Saladores, E. Lambert, A. O. Hudson, and T. Leustek Localization of Members of the {gamma}-Glutamyl Transpeptidase Family Identifies Sites of Glutathione and Glutathione S-Conjugate Hydrolysis Plant Physiology, August 1, 2007; 144(4): 1715 - 1732. [Abstract] [Full Text] [PDF]

				M. Hothorn, A. Wachter, R. Gromes, T. Stuwe, T. Rausch, and K. Scheffzek Structural Basis for the Redox Control of Plant Glutamate Cysteine Ligase J. Biol. Chem., September 15, 2006; 281(37): 27557 - 27565. [Abstract] [Full Text] [PDF]

				J. Hancock, R. Desikan, J. Harrison, J. Bright, R. Hooley, and S. Neill Doing the unexpected: proteins involved in hydrogen peroxide perception J. Exp. Bot., May 1, 2006; 57(8): 1711 - 1718. [Abstract] [Full Text] [PDF]