title: Ischemia/reperfusion-induced myosin light chain 1 phosphorylation increases its degradation by matrix metalloproteinase 2.
abstract: Degradation of myosin light chain 1 (MLC1) by matrix metalloproteinase 2 (MMP-2) during myocardial ischemia/reperfusion (I/R) has been demonstrated. However, the exact mechanisms controlling this process remain unknown. I/R increases the phosphorylation of MLC1, but the consequences of this modification are not known. We hypothesized that phosphorylation of MLC1 plays an important role in its degradation by MMP-2. To examine this, isolated perfused rat hearts were subjected to 20 min global ischemia followed by 30 min of aerobic reperfusion. I/R increased phosphorylation of MLC1 (as measured by mass spectrometry). When hearts were subjected to I/R in the presence of ML-7 (a myosin light-chain kinase inhibitor) or doxycycline (an MMP inhibitor), improved recovery of contractile function was observed compared to aerobic controls, and MLC1 was protected from degradation. Enzyme kinetic studies revealed an increased affinity of MMP-2 for the phosphorylated form of MLC1 compared to non-phosphorylated MLC1. We conclude that MLC1 phosphorylation is an important mechanism controlling the intracellular action of MMP-2 and promoting degradation of MLC1. These results further support previous findings implicating post-translational modifications of contractile proteins as a key factor in the pathology of cardiac dysfunction during and following ischemia.
authors.affiliation: Department of Pharmacology, University of Saskatchewan, Saskatoon, SK, Canada.
authors.forename: Virgilio J J
authors.initials: VJ
authors.lastname: Cadete
authors.forename: Jolanta
authors.initials: J
authors.lastname: Sawicka
authors.forename: Jagdip S
authors.initials: JS
authors.lastname: Jaswal
authors.forename: Gary D
authors.initials: GD
authors.lastname: Lopaschuk
authors.forename: Richard
authors.initials: R
authors.lastname: Schulz
authors.forename: Danuta
authors.initials: D
authors.lastname: Szczesna-Cordary
authors.forename: Grzegorz
authors.initials: G
authors.lastname: Sawicki
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D004318:Doxycycline / Q000494:pharmacology; D015180:Electrophoresis, Gel, Two-Dimensional; D015151:Immunoblotting; D007700:Kinetics; D008297:Male; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D017202:Myocardial Ischemia / Q000503:physiopathology*; D015428:Myocardial Reperfusion Injury / Q000503:physiopathology*; D018994:Myosin Light Chains / Q000378:metabolism*; D009219:Myosin-Light-Chain Kinase / Q000378:metabolism; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D059748:Proteolysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000900:Anti-Bacterial Agents; C425069:MLC1 protein, human; D008565:Membrane Proteins; D018994:Myosin Light Chains; C076659:myosin light chain I; D009219:Myosin-Light-Chain Kinase; D020778:Matrix Metalloproteinase 2; D004318:Doxycycline