Abstract
Left ventricular remodeling post-myocardial infarction (MI) involves a multitude of mechanisms that regulate the repair response. Matrix metalloproteinases (MMPs) are a major family of proteolytic enzymes that coordinate extracellular matrix turnover. MMP-7 or MMP-9 deletion attenuate adverse remodeling post-MI, but the mechanisms have not been fully clarified. Both MMP-7 and MMP-9 have a large number of known in vitro substrates, but in vivo substrates for these two MMPs in the myocardial infarction setting are incompletely identified. Advances in proteomic techniques have enabled comprehensive profiling of protein expression in cells and tissue. In this chapter, we describe a protocol for the proteomic analysis of in vivo candidate MMP substrates in the post-MI left ventricle using two-dimensional electrophoresis, liquid chromatography coupled with tandem mass spectrometry, and immunoblotting.
Andriy Yabluchanskiy and Yaojun Li have contributed equally to this chapter.
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References
Dixon JA, Spinale FG (2011) Myocardial remodeling: cellular and extracellular events and targets. Annu Rev Physiol 73:47–68
Lindsey ML, Weintraub ST, Lange RA (2012) Using extracellular matrix proteomics to understand left ventricular remodeling. Circ Cardiovasc Genet 5:o1–7
Gajarsa JJ, Kloner RA (2011) Left ventricular remodeling in the post-infarction heart: a review of cellular, molecular mechanisms, and therapeutic modalities. Heart Fail Rev 16:13–21
Page-McCaw A, Ewald AJ, Werb Z (2007) Matrix metalloproteinases and the regulation of tissue remodelling. Nat Rev Mol Cell Biol 8:221–233
Kandalam V, Basu R, Abraham T et al (2010) Early activation of matrix metalloproteinases underlies the exacerbated systolic and diastolic dysfunction in mice lacking TIMP3 following myocardial infarction. Am J Physiol Heart Circ Physiol 299:H1012–1023
Lindsey ML, Zamilpa R (2012) Temporal and spatial expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases following myocardial infarction. Cardiovasc Ther 30:31–41
Lindsey ML, Escobar GP, Dobrucki LW et al (2006) Matrix metalloproteinase-9 gene deletion facilitates angiogenesis after myocardial infarction. Am J Physiol Heart Circ Physiol 290:H232–239
Lindsey ML, Escobar GP, Mukherjee R et al (2006) Matrix metalloproteinase-7 affects connexin-43 levels, electrical conduction, and survival after myocardial infarction. Circulation 113:2919–2928
Agnihotri R, Crawford HC, Haro H et al (2001) Osteopontin, a novel substrate for matrix metalloproteinase-3 (stromelysin-1) and matrix metalloproteinase-7 (matrilysin). J Biol Chem 276:28261–28267
Siri A, Knauper V, Veirana N et al (1995) Different susceptibility of small and large human tenascin-C isoforms to degradation by matrix metalloproteinases. J Biol Chem 270:8650–8654
von Bredow DC, Nagle RB, Bowden GT et al (1995) Degradation of fibronectin fibrils by matrilysin and characterization of the degradation products. Exp Cell Res 221:83–91
Imai K, Yokohama Y, Nakanishi I et al (1995) Matrix metalloproteinase 7 (matrilysin) from human rectal carcinoma cells. Activation of the precursor, interaction with other matrix metalloproteinases and enzymic properties. J Biol Chem 270:6691–6697
Lee KH, Choi EY, Hyun MS et al (2007) Association of extracellular cleavage of E-cadherin mediated by MMP-7 with HGF-induced in vitro invasion in human stomach cancer cells. Eur Surg Res 39:208–215
Belaaouaj AA, Li A, Wun TC et al (2000) Matrix metalloproteinases cleave tissue factor pathway inhibitor. Effects on coagulation. J Biol Chem 275:27123–27128
Crabbe T, Smith B, O'Connell J et al (1994) Human progelatinase A can be activated by matrilysin. FEBS Lett 345:14–16
Trocme C, Gaudin P, Berthier S et al (1998) Human B lymphocytes synthesize the 92-kDa gelatinase, matrix metalloproteinase-9. J Biol Chem 273:20677–20684
Birkedal-Hansen H (1995) Proteolytic remodeling of extracellular matrix. Curr Opin Cell Biol 7:728–735
Visse R, Nagase H (2003) Matrix metalloproteinases and tissue inhibitors of metalloproteinases: structure, function, and biochemistry. Circ Res 92:827–839
Tan RJ, Liu Y (2012) Matrix metalloproteinases in kidney homeostasis and diseases. Am J Physiol Ren Physiol 302:F1351–1361
Yu Q, Stamenkovic I (2000) Cell surface-localized matrix metalloproteinase-9 proteolytically activates TGF-beta and promotes tumor invasion and angiogenesis. Genes Dev 14:163–176
McCawley LJ, Matrisian LM (2001) Matrix metalloproteinases: they're not just for matrix anymore! Curr Opin Cell Biol 13:534–540
Zamilpa R, Lopez EF, Chiao YA et al (2010) Proteomic analysis identifies in vivo candidate matrix metalloproteinase-9 substrates in the left ventricle post-myocardial infarction. Proteomics 10:2214–2223
Chiao YA, Zamilpa R, Lopez EF et al (2010) In vivo matrix metalloproteinase-7 substrates identified in the left ventricle post-myocardial infarction using proteomics. J Proteome Res 9:2649–2657
Haro H, Crawford HC, Fingleton B et al (2000) Matrix metalloproteinase-7-dependent release of tumor necrosis factor-alpha in a model of herniated disc resorption. J Clin Invest 105:143–150
Acuff HB, Carter KJ, Fingleton B et al (2006) Matrix metalloproteinase-9 from bone marrow-derived cells contributes to survival but not growth of tumor cells in the lung microenvironment. Cancer Res 66:259–266
Acknowledgments
We acknowledge support from NIH/NHLBI HHSN 268201000036C (N01-HV-00244) for the San Antonio Cardiovascular Proteomics Center and R01 HL075360, the Max and Minnie Tomerlin Voelcker Fund, and the Veteran’s Administration (Merit) to M.L.L. Mass spectrometry analyses were conducted in the UTHSCSA Institutional Mass Spectrometry Laboratory.
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Yabluchanskiy, A., Li, Y., de Castro Brás, L.E., Hakala, K., Weintraub, S.T., Lindsey, M.L. (2013). Proteomic Analysis of the Left Ventricle Post-myocardial Infarction to Identify In Vivo Candidate Matrix Metalloproteinase Substrates. In: Baudino, T. (eds) Cell-Cell Interactions. Methods in Molecular Biology, vol 1066. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-604-7_16
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DOI: https://doi.org/10.1007/978-1-62703-604-7_16
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