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Peroxynitrite‐induced changes in 72kDa matrix metalloproteinase‐2 activity are further regulated by its phosphorylation status
Author(s) -
Ferreira Anna Laura Bechara Jacob,
Schulz Richard
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.1096.2
Subject(s) - peroxynitrite , matrix metalloproteinase , phosphorylation , chemistry , microbiology and biotechnology , matrix metalloproteinase 9 , matrix (chemical analysis) , biochemistry , biology , enzyme , chromatography , superoxide
Matrix metalloproteinase‐2 (MMP‐2) is a key protease in several oxidative stress related pathologies. 72kDa MMP‐2 activity is modulated by peroxynitrite (ONOO − ), or phosphorylation, however, the effect of these together has not been tested. The activity of human recombinant 72kDa MMP‐2 following various in vitro treatments was measured by gelatin zymography. ONOO − treatment (15min, 37°C) resulted in concentration‐dependent changes in MMP‐2 activity, with 0.3–10μM increasing and 100μM attenuating its activity. Dephosphorylation of MMP‐2 with alkaline phosphatase (20min, 37°C) significantly increased its activity, either with or without ONOO − . Glutathione (GSH, 30μM) without or with 100μM ONOO − , decreased MMP‐2 activity. Treatment of MMP‐2 with 0.3μM ONOO − in the presence of 30μM glutathione increased its activity and even moreso its dephosphorylated form. GSH did not significantly affect MMP‐2 activity with 1–10μM ONOO − . With 100μM ONOO − , dephosphorylation partially protected MMP‐2 from the attenuation of its activity. These results suggest that ONOO − activation (at low μM) and inactivation (at high μM) of 72kDa MMP‐2, in the presence or absence of glutathione, is further modulated by its phosphorylation status. This may be relevant to pathophysiological conditions associated with increased oxidative stress and intracellular activation of MMP‐2. Grant support from the CIHR.