Inactivation of tissue inhibitor of metalloproteinase-1 by peroxynitrite.
Peroxynitrite (ONOO-) has recently been implicated in connective tissue destruction in vivo. We have studied the effect of ONOO- on the activity of tissue inhibitor of metalloproteinase-1 (TIMP-1) in vitro. The inactivation of TIMP-1 by ONOO- was dose dependent with 50 microM ONOO- reducing the inhibitory activity of TIMP-1 towards gelatinase-A by 50%. High concentrations of ONOO- (500 microM-5 mM) caused protein fragmentation whilst lower concentrations (<250 microM) inactivated TIMP-1 without altering the molecular weight. Inactivation could be blocked by ONOO- scavengers but not by hydroxyl radical scavengers. Our results show that ONOO- is capable of inactivating TIMP-1, a process which could potentiate metalloproteinase-mediated tissue breakdown.
Frears
E R
ER
The inflammation Research Group, London Hospital Medical College, University of London, UK.
Zhang
Z
Z
Blake
D R
DR
O'Connell
J P
JP
Winyard
P G
PG
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D016166:Free Radical Scavengers / Q000494:pharmacology; D006023:Glycoproteins / Q000037:antagonists & inhibitors* / Q000302:isolation & purification / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D019278:Matrix Metalloproteinase 3; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008969:Molecular Sequence Data; D009566:Nitrates / Q000138:chemical synthesis / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000494:pharmacology; D013379:Substrate Specificity; D019714:Tissue Inhibitor of Metalloproteinases
D000596:Amino Acids; D016166:Free Radical Scavengers; D006023:Glycoproteins; D009566:Nitrates; D011994:Recombinant Proteins; D019714:Tissue Inhibitor of Metalloproteinases; C039642:peroxynitric acid; D008666:Metalloendopeptidases; D019278:Matrix Metalloproteinase 3