HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 279, Issue 40, 41352-41360, October 1, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: 279/40/41352    most recent M403827200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal 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 Google Scholar Google Scholar Articles by Brennan, J. P. Articles by Eaton, P. Search for Related Content PubMed PubMed Citation Articles by Brennan, J. P. Articles by Eaton, P. Social Bookmarking                      What's this?

Detection and Mapping of Widespread Intermolecular Protein Disulfide Formation during Cardiac Oxidative Stress Using Proteomics with Diagonal Electrophoresis^*

Jonathan P. Brennan, Robin Wait, Shajna Begum, James R. Bell, Michael J. Dunn¶, and Philip Eaton||

From the Department of Cardiology, Cardiovascular Division, King's College London, The Rayne Institute, St. Thomas' Hospital, London SE1 7EH, United Kingdom, the Kennedy Institute of Rheumatology Division, Faculty of Medicine, Imperial College London, 1 Aspenlea Road, Hammersmith, London, W6 8LH, United Kingdom, and the ^¶Department of Neuroscience, Institute of Psychiatry, King's College London, London SE5 8AF, United Kingdom

Regulation of protein function by reversible cysteine-targeted oxidation can be achieved by multiple mechanisms, such as S-glutathiolation, S-nitrosylation, sulfenic acid, sulfinic acid, and sulfenyl amide formation, as well as intramolecular disulfide bonding of vicinal thiols. Another cysteine oxidation state with regulatory potential involves the formation of intermolecular protein disulfides. We utilized two-dimensional sequential non-reducing/reducing SDS-PAGE (diagonal electrophoresis) to investigate intermolecular protein disulfide formation in adult cardiac myocytes subjected to a series of interventions (hydrogen peroxide, S-nitroso-N-acetylpenicillamine, doxorubicin, simulated ischemia, or metabolic inhibition) that alter the redox status of the cell. More detailed experiments were undertaken with the thiol-specific oxidant diamide (5 mM), a concentration that induces a mild non-injurious oxidative stress. This increase in cellular oxidation potential caused global intermolecular protein disulfide formation in cytosolic, membrane, and myofilament/cytoskeletal compartments. A large number of proteins that undergo these associations were identified using liquid chromatography-mass spectrometry/mass spectrometry. These associations, which involve metabolic and antioxidant enzymes, structural proteins, signaling molecules, and molecular chaperones, were confirmed by assessing "shifts" on non-reducing immunoblots. The observation of widespread protein-protein disulfides indicates that these oxidative associations are likely to be fundamental in how cells respond to redox changes.

Received for publication, April 6, 2004 , and in revised form, July 26, 2004.

^* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^|| To whom correspondence should be addressed: Dept. of Cardiology, The Rayne Institute, St. Thomas' Hospital, London, SE1 7EH. Tel.: 44-020-7188-0969; Fax: 44-020-7188-0970; E-mail: philip.eaton{at}kcl.ac.uk.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				E. Schroder, J. P. Brennan, and P. Eaton Cardiac peroxiredoxins undergo complex modifications during cardiac oxidant stress Am J Physiol Heart Circ Physiol, July 1, 2008; 295(1): H425 - H433. [Abstract] [Full Text] [PDF]

				M. Z. de Pina, H. Vazquez-Meza, J. P. Pardo, J. L. Rendon, R. Villalobos-Molina, H. Riveros-Rosas, and E. Pina Signaling the Signal, Cyclic AMP-dependent Protein Kinase Inhibition by Insulin-formed H2O2 and Reactivation by Thioredoxin J. Biol. Chem., May 2, 2008; 283(18): 12373 - 12386. [Abstract] [Full Text] [PDF]

				J. R. Burgoyne, M. Madhani, F. Cuello, R. L. Charles, J. P. Brennan, E. Schroder, D. D. Browning, and P. Eaton Cysteine Redox Sensor in PKGIa Enables Oxidant-Induced Activation Science, September 7, 2007; 317(5843): 1393 - 1397. [Abstract] [Full Text] [PDF]

				R. L. Charles, E. Schroder, G. May, P. Free, P. R. J. Gaffney, R. Wait, S. Begum, R. J. Heads, and P. Eaton Protein Sulfenation as a Redox Sensor: Proteomics Studies Using a Novel Biotinylated Dimedone Analogue Mol. Cell. Proteomics, September 1, 2007; 6(9): 1473 - 1484. [Abstract] [Full Text] [PDF]

				J. P. Brennan, S. C. Bardswell, J. R. Burgoyne, W. Fuller, E. Schroder, R. Wait, S. Begum, J. C. Kentish, and P. Eaton Oxidant-induced Activation of Type I Protein Kinase A Is Mediated by RI Subunit Interprotein Disulfide Bond Formation J. Biol. Chem., August 4, 2006; 281(31): 21827 - 21836. [Abstract] [Full Text] [PDF]

				N. Le Moan, G. Clement, S. Le Maout, F. Tacnet, and M. B. Toledano The Saccharomyces cerevisiae Proteome of Oxidized Protein Thiols: CONTRASTED FUNCTIONS FOR THE THIOREDOXIN AND GLUTATHIONE PATHWAYS J. Biol. Chem., April 14, 2006; 281(15): 10420 - 10430. [Abstract] [Full Text] [PDF]

				M. Canton, A. Skyschally, R. Menabo, K. Boengler, P. Gres, R. Schulz, M. Haude, R. Erbel, F. Di Lisa, and G. Heusch Oxidative modification of tropomyosin and myocardial dysfunction following coronary microembolization Eur. Heart J., April 1, 2006; 27(7): 875 - 881. [Abstract] [Full Text] [PDF]

				T. R. Moopanar and D. G. Allen The activity-induced reduction of myofibrillar Ca2+ sensitivity in mouse skeletal muscle is reversed by dithiothreitol J. Physiol., February 15, 2006; 571(1): 191 - 200. [Abstract] [Full Text] [PDF]

				K. M. Humphries, M. S. Deal, and S. S. Taylor Enhanced Dephosphorylation of cAMP-dependent Protein Kinase by Oxidation and Thiol Modification J. Biol. Chem., January 28, 2005; 280(4): 2750 - 2758. [Abstract] [Full Text] [PDF]