Activation of Matrix Metalloproteinases by Peroxynitrite-induced Protein S-Glutathiolation via Disulfide S-Oxide Formation

Abstract

Oxidative stress may cause tissue injury through activation of the precursors of matrix metalloproteinase (proMMPs). In this study, we observed glutathione (GSH)-dependent proMMP activation induced by peroxynitrite, a potent oxidizing agent formed during inflammatory processes. Peroxynitrite strongly activated all three types of purified human proMMPs (proMMP-1, -8, and -9) in the presence of similar concentrations of GSH. Of the potential reaction products between peroxynitrite and GSH, only S-nitroglutathione (GSNO 2 ) caused proMMP activation. Extensive S-glutathiolation of the proMMP protein occurred during activation of proMMP by peroxynitrite and GSH, as shown by radiolabeling studies with [ S]GSH or [3 H]GSH. Evidence of appreciable S-glutathiolation persisted even after dithiothreitol and proteindenaturing treatment, however, suggesting that some Sglutathiolation did not occur through formation of simple mixed disulfide. Matrix-assisted laser-desorption ionization-time-of-flight mass spectrometry indicated that not only peroxynitrite plus GSH but also synthetic GSNO 2 produced dithiothreitol-resistant S-glutathiolation of the synthetic peptide PRCGVPD, which is a well conserved Cys-containing sequence of the propeptide autoinhibitory domain of proMMPs. PRCGVPD S-glutathiolation is presumed to be formed through glutathione disulfide Soxide (GS(O)SR), based on the m/z 1064. Our results illustrate a unique mechanism of oxidative proMMP activation and oxidative tissue injury during inflammation.

Cited2 modulates TGF-β-mediated upregulation of MMP9

“…Full-length MMP9 is the major form of MMP9 in cancer tissues (Fridman et al, 2003) and in MDA-MB-231, an invasive breast cancer cell line. Several studies have suggested that without the participation of proteolytic enzymes and removal of the inhibitory peptide, full-length MMP9 can be activated by binding to the substrates to cause conformational changes or oxidative modification of the cysteine sidechain thiol groups, in turn allowing zinc ion to become available for the catalytic function (Okamoto et al, 2001;Bannikov et al, 2002;Gu et al, 2002). Transforming growth factor-b is a potent growth inhibitor of normal epithelial cells, but also fosters tumor formation during later stages of tumorigenesis (Wakefield and Roberts, 2002).…”

Regulation of Annexin A2 by Reversible Glutathionylation

“…Although the majority of studies have reported an inhibition of enzyme activity by glutathionylation, it appears that the position of the cysteine undergoing modification is important in determining the effect of glutathionylation. For instance, human immunodeficiency virus type 1 protease can be inhibited or stabilized by glutathionylation depending on which cysteine is involved (39), whereas matrix metalloproteinases are activated upon glutathionylation of the autoinhibitory domain (40).…”

Potent Metalloporphyrin Peroxynitrite Decomposition Catalyst Protects Against the Development of Doxorubicin-Induced Cardiac Dysfunction

“…The activation of proMMPs is triggered by peroxynitrite generation via an extensive S-glutathiolation reaction. 29 By inhibiting this reaction, peroxynitrite decomposition catalysts may reduce MMP activation. In addition to direct oxidation, peroxidation, and nitration reactions and MMP activation, likely additional downstream cytotoxic mechanisms elicited by peroxynitrite during DOX-induced cardiac injury include DNA injury and activation of the nuclear enzyme poly(ADP-ribose) polymerase, as well as the inhibition of myofibrillar CK.…”