title: A sequential extraction methodology for cardiac extracellular matrix prior to proteomics analysis.
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.
authors.affiliation: King's British Heart Foundation Centre, London, UK.
authors.forename: Javier
authors.initials: J
authors.lastname: Barallobre-Barreiro
authors.forename: Athanasios
authors.initials: A
authors.lastname: Didangelos
authors.forename: Xiaoke
authors.initials: X
authors.lastname: Yin
authors.forename: Nieves
authors.initials: N
authors.lastname: Doménech
authors.forename: Manuel
authors.initials: M
authors.lastname: Mayr
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid; D005109:Extracellular Matrix / Q000737:chemistry* / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D005355:Fibrosis; D019791:Guanidine; D006801:Humans; D059625:Liquid-Liquid Extraction / Q000379:methods*; D015428:Myocardial Reperfusion Injury / Q000378:metabolism* / Q000473:pathology; D009206:Myocardium / Q000737:chemistry* / Q000378:metabolism / Q000473:pathology; D010446:Peptide Fragments / Q000737:chemistry* / Q000378:metabolism; D010449:Peptide Mapping; D059748:Proteolysis; D040901:Proteomics; D012965:Sodium Chloride; D012967:Sodium Dodecyl Sulfate; D013552:Swine; D053719:Tandem Mass Spectrometry
chemical_list: D016326:Extracellular Matrix Proteins; D010446:Peptide Fragments; D012967:Sodium Dodecyl Sulfate; D012965:Sodium Chloride; D019791:Guanidine