title: Inhibitory effects of caspase inhibitors on the activity of matrix metalloproteinase-2.
abstract: Matrix metalloproteinase (MMP)-2, a zinc-dependent endopeptidase, plays a detrimental role in several diseases including ischemia and reperfusion (I/R) injury of the heart. Caspases are a group of cysteine-dependent, aspartate-directed proteases which regulate cellular apoptosis. Interestingly, protective effects of caspase inhibitors independent of apoptosis have been shown in I/R injury of the heart. The cardioprotective actions of both these classes of protease inhibitors led us to hypothesize that caspase inhibitors may also reduce MMP-2 activity. Five known caspase inhibitors (Z-IE(OMe)TD(OMe)-fmk, Ac-DEVD-CHO, Ac-LEHD-cmk, Z-VAD-fmk and Ac-YVAD-cmk) were tested for their possible inhibitory effects on MMP-2 activity in comparison to the MMP inhibitors ONO-4817 and ARP-100 (which themselves were unable to inhibit caspase-3 activity). MMP-2 activity was assessed by an in vitro troponin I (TnI) proteolysis assay and a quantitative kinetic fluorescence assay using a fluorogenic peptide substrate (OmniMMP). TnI proteolysis was also measured by western blot in neonatal cardiomyocytes subjected to hypoxia-reoxygenation injury. Using human recombinant MMP-2 and TnI as its substrate, the caspase inhibitors, in comparison with ONO-4817, significantly inhibited MMP-2-mediated TnI degradation in a concentration-dependent manner. The kinetic assay using OmniMMP revealed that these caspase inhibitors blocked MMP-2 activity in a concentration-dependent manner with similar IC50 values. TnI degradation in neonatal cardiomyocytes was enhanced following hypoxia-reoxygenation and this was blocked by ARP-100 and Ac-LEHD-cmk. Inhibition of MMP-2 activity is an additional pharmacological action which contributes to the protective effects of some caspase inhibitors.
authors.affiliation: Department of Pharmacology, Cardiovascular Research Centre, Mazankowski Alberta Heart Institute, University of Alberta, Edmonton, AB, Canada T6G 2S2.
authors.forename: M M
authors.initials: MM
authors.lastname: Castro
authors.forename: J
authors.initials: J
authors.lastname: Fuah
authors.forename: M
authors.initials: M
authors.lastname: Ali
authors.forename: M
authors.initials: M
authors.lastname: Sung
authors.forename: J
authors.initials: J
authors.lastname: Schulz
authors.forename: M Y
authors.initials: MY
authors.lastname: Kondo
authors.forename: X
authors.initials: X
authors.lastname: Fan
authors.forename: A
authors.initials: A
authors.lastname: Holt
authors.forename: R
authors.initials: R
authors.lastname: Schulz
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D053148:Caspase 3 / Q000378:metabolism; D061945:Caspase Inhibitors / Q000494:pharmacology*; D015687:Cell Hypoxia; D002478:Cells, Cultured; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D061965:Matrix Metalloproteinase Inhibitors / Q000494:pharmacology*; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000201:enzymology; D009842:Oligopeptides / Q000494:pharmacology; D010647:Phenyl Ethers / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013450:Sulfones / Q000494:pharmacology; D019210:Troponin I / Q000378:metabolism
chemical_list: D061945:Caspase Inhibitors; D006877:Hydroxamic Acids; D061965:Matrix Metalloproteinase Inhibitors; C583722:N-hydroxy-2-((4-phenylphenyl)sulfonylpropan-2-yloxyamino)acetamide; C414368:N-hydroxy-5-ethoxymethyloxy-2-methyl-4-(4-phenoxybenzoyl)aminopentanamide; D009842:Oligopeptides; D010647:Phenyl Ethers; D013450:Sulfones; D019210:Troponin I; D053148:Caspase 3; D020778:Matrix Metalloproteinase 2
keywords: Caspase inhibitors; Caspases; Hypoxia-reoxygenation; I/R; MMPs; Matrix metalloproteinase-2; TnI; Troponin I; ischemia and reperfusion; matrix metalloproteinases; troponin I