Activation of matrix metalloproteinases by peroxynitrite-induced protein S-glutathiolation via disulfide S-oxide formation.
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 [(35)S]GSH or [(3)H]GSH. Evidence of appreciable S-glutathiolation persisted even after dithiothreitol and protein-denaturing treatment, however, suggesting that some S-glutathiolation 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 S-oxide (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.
Okamoto
T
T
Department of Microbiology, Kumamoto University School of Medicine, Kumamoto 860-0811, Japan.
Akaike
T
T
Sawa
T
T
Miyamoto
Y
Y
van der Vliet
A
A
Maeda
H
H
D000595:Amino Acid Sequence; D001665:Binding Sites; D017364:Collagenases / Q000378:metabolism*; D004220:Disulfides / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D004792:Enzyme Precursors / Q000378:metabolism*; D005978:Glutathione / Q000031:analogs & derivatives / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D020780:Matrix Metalloproteinase 9; D020782:Matrix Metalloproteinases / Q000378:metabolism*; D009566:Nitrates / Q000378:metabolism / Q000494:pharmacology*; D009603:Nitroso Compounds / Q000494:pharmacology; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000737:chemistry; D011499:Protein Processing, Post-Translational; D026422:S-Nitrosoglutathione; D012710:Serum Albumin, Bovine / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013462:Sulfur Radioisotopes; D014316:Tritium
D004220:Disulfides; D004792:Enzyme Precursors; D009566:Nitrates; D009603:Nitroso Compounds; D010446:Peptide Fragments; D013462:Sulfur Radioisotopes; D014316:Tritium; C039642:peroxynitric acid; D012710:Serum Albumin, Bovine; D026422:S-Nitrosoglutathione; D017364:Collagenases; D020782:Matrix Metalloproteinases; C120487:pro-matrix metalloproteinase 9; D020780:Matrix Metalloproteinase 9; D005978:Glutathione