Oxidative Modification and Inactivation of the Proteasome during Coronary Occlusion/Reperfusion

Anne Laure Bulteau, Kathleen C. Lundberg, Kenneth M. Humphries, Hesham A. Sadek, Pamela A. Szweda, Bertrand Friguet, Luke I. Szweda

Research output: Contribution to journalArticle

300 Citations (Scopus)

Abstract

Restoration of blood flow to ischemic myocardial tissue results in an increase in the production of oxygen radicals. Highly reactive, free radical species have the potential to damage cellular components. Clearly, maintenance of cellular viability is dependent, in part, on the removal of altered protein. The proteasome is a major intracellular proteolytic system which degrades oxidized and ubiquitinated forms of protein. Utilizing an in vivo rat model, we demonstrate that coronary occlusion/reperfusion resulted in declines in chymotrypsin-like, peptidylglutamyl-peptide hydrolase, and trypsin-like activities of the proteasome as assayed in cytosolic extracts. Analysis of purified 20 S proteasome revealed that declines in peptidase activities were accompanied by oxidative modification of the protein. We provide conclusive evidence that, upon coronary occlusion/ reperfusion, the lipid peroxidation product 4-hydroxy-2-nonenal selectively modifies 20 S proteasome α-like subunits iota, C3, and an isoform of XAPC7. Occlusion/reperfusion-induced declines in trypsin-like activity were largely preserved upon proteasome purification. In contrast, loss in chymotrypsin-like and peptidylglutamyl-peptide hydrolase activities observed in cytosolic extracts were not evident upon purification. Thus, decreases in proteasome activity are likely due to both direct oxidative modification of the enzyme and inhibition of fluorogenic peptide hydrolysis by endogenous cytosolic inhibitory protein(s) and/or substrate(s). Along with inhibition of the proteasome, increases in cytosolic levels of oxidized and ubiquitinated protein(s) were observed. Taken together, our findings provide insight into potential mechanisms of coronary occlusion/reperfusion-induced proteasome inactivation and cellular consequences of these events.

Original languageEnglish (US)
Pages (from-to)30057-30063
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number32
DOIs
StatePublished - Aug 10 2001

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Myocardial Reperfusion
Coronary Occlusion
Proteasome Endopeptidase Complex
Ubiquitinated Proteins
Peptide Hydrolases
Chymotrypsin
Trypsin
Purification
Proteins
Lipid Peroxidation
Restoration
Reperfusion
Free Radicals
Rats
Hydrolysis
Reactive Oxygen Species
Protein Isoforms
Blood
Maintenance
Tissue

ASJC Scopus subject areas

  • Biochemistry

Cite this

Oxidative Modification and Inactivation of the Proteasome during Coronary Occlusion/Reperfusion. / Bulteau, Anne Laure; Lundberg, Kathleen C.; Humphries, Kenneth M.; Sadek, Hesham A.; Szweda, Pamela A.; Friguet, Bertrand; Szweda, Luke I.

In: Journal of Biological Chemistry, Vol. 276, No. 32, 10.08.2001, p. 30057-30063.

Research output: Contribution to journalArticle

Bulteau, Anne Laure ; Lundberg, Kathleen C. ; Humphries, Kenneth M. ; Sadek, Hesham A. ; Szweda, Pamela A. ; Friguet, Bertrand ; Szweda, Luke I. / Oxidative Modification and Inactivation of the Proteasome during Coronary Occlusion/Reperfusion. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 32. pp. 30057-30063.
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