Activation and modulation of 72kDa matrix metalloproteinase-2 by peroxynitrite and glutathione.
Matrix metalloproteinase-2 (MMP-2) has emerged as a key protease in various pathologies associated with oxidative stress, including myocardial ischemia-reperfusion, heart failure or inflammation. Peroxynitrite (ONOO(-)), an important effector of oxidative stress, was reported to activate some full length MMP zymogens, particularly in the presence of glutathione (GSH), but whether this occurs for MMP-2 is unknown. Treating MMP-2 zymogen with ONOO(-) resulted in a concentration-dependent regulation of MMP-2, with 0.3-1 microM ONOO(-) increasing and 30-100 microM ONOO(-) attenuating enzyme activity. The enzyme's V(max) was also significantly increased by 1 microM ONOO(-). Comparable responses to ONOO(-) treatment were observed using the intracellular target of MMP-2, troponin I (TnI). GSH at 100 microM attenuated the effects of ONOO(-) on MMP-2. Mass spectrometry revealed that ONOO(-) can oxidize and, in the presence of GSH, S-glutathiolate the MMP-2 zymogen or a synthetic peptide containing the cysteine-switch motif in the enzyme's autoinhibitory domain. These results suggest that ONOO(-) and GSH can modulate the activity of 72 kDa MMP-2 by modifying the cysteine residue in the autoinhibitory domain of the zymogen, a process that may be relevant to pathophysiological conditions associated with increased oxidative stress.
Viappiani
Serena
S
Cardiovascular Research Group, Departments of Pediatrics and Pharmacology, University of Alberta, Edmonton, AB T6G2S2, Canada.
Nicolescu
Adrian C
AC
Holt
Andrew
A
Sawicki
Grzegorz
G
Crawford
Bryan D
BD
León
Hernando
H
van Mulligen
Tyler
T
Schulz
Richard
R
D000595:Amino Acid Sequence; D004789:Enzyme Activation; D005978:Glutathione / Q000494:pharmacology*; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D018384:Oxidative Stress; D030421:Peroxynitrous Acid / Q000494:pharmacology*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry
D030421:Peroxynitrous Acid; D020778:Matrix Metalloproteinase 2; D005978:Glutathione