Abstract
Cardiac fibrosis is characterized by excessive deposition of extracellular matrix (ECM) and is a common complication of various cardiovascular diseases. However, little is known about proteins in the cardiac extracellular space. Proteomics analysis of cardiac ECM can be challenging due to the presence of more abundant intracellular proteins, the low degree of solubility of integral ECM proteins, and the presence of abundant posttranslational modifications. Here we describe an extraction methodology based on tissue decellularization, which allows the biochemical subfractionation of extracellular proteins in cardiac tissue. These relatively low-complexity protein fractions are suitable for analysis by gel-LC-MS/MS and other proteomics techniques.
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References
Agnetti G, Husberg C, Van Eyk JE (2011) Divide and conquer: the application of organelle proteomics to heart failure. Circ Res 108:512–526
Matsui Y, Morimoto J, Uede T (2010) Role of matricellular proteins in cardiac tissue remodeling after myocardial infarction. World J Biol Chem 1:69–80
Bowers SL, Banerjee I, Baudino TA (2010) The extracellular matrix: at the center of it all. J Mol Cell Cardiol 48:474–482
Vogel KG, Peters JA (2001) Isolation of proteoglycans from tendon. Methods Mol Biol 171:9–17
Barallobre-Barreiro J, Didangelos A, Schoendube FA, Drozdov I, Yin X, Fernández-Caggiano M, Willeit P, Puntmann VO, Aldama-López G, Shah AM, Doménech N, Mayr M (2012) Proteomics analysis of cardiac extracellular matrix remodeling in a porcine model of ischemia-reperfusion injury. Circulation 125:789–802
Didangelos A, Yin X, Mandal K, Baumert M, Jahangiri M, Mayr M (2010) Proteomics characterization of extracellular space components in the human aorta. Mol Cell Proteomics 9:2048–2062
Didangelos A, Yin X, Mayr M (2012) Method for protein subfractionation of cardiovascular tissues before DIGE analysis. Methods Mol Biol 854:287–297
Mason RM, Mayes RW (1973) Extraction of cartilage protein-polysaccharides with inorganic salt solutions. Biochem J 131:535–540
Stoscheck CM (1990) Quantitation of protein. Methods Enzymol 182:50–68
Stanton H, Fosang AJ (2002) Matrix metalloproteinase are active following guanidine hydrochloride extraction of cartilage: generation of DIPEN neoepitope during dialysis. Matrix Biol 21:425–428
Didangelos A, Yin X, Mandal K, Saje A, Smith A, Xu Q, Jahangiri M, Mayr M (2011) Extracellular matrix composition and remodeling in human abdominal aortic aneurysms: a proteomics approach. Mol Cell Proteomics 10:M111.008128
Shevchenko A, Tomas H, Havlis J, Olsen JV, Mann M (2006) In-gel digestion for mass spectrometric characterization of proteins and proteomes. Nat Protoc 1:2856–2860
Yin X, Cuello F, Mayr U, Hao Z, Hornshaw M, Ehler E, Avkiran M, Mayr M (2010) Proteomics analysis of the cardiac myofilament subproteome reveals dynamic alterations in phosphatase subunit distribution. Mol Cell Proteomics 9:497–509
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Barallobre-Barreiro, J., Didangelos, A., Yin, X., Doménech, N., Mayr, M. (2013). A Sequential Extraction Methodology for Cardiac Extracellular Matrix Prior to Proteomics Analysis. In: Vivanco, F. (eds) Heart Proteomics. Methods in Molecular Biology, vol 1005. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-386-2_17
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DOI: https://doi.org/10.1007/978-1-62703-386-2_17
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-385-5
Online ISBN: 978-1-62703-386-2
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