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蛋白质组学分析前用于心脏细胞外基质的顺序提取方法。

A sequential extraction methodology for cardiac extracellular matrix prior to proteomics analysis.

作者信息

Barallobre-Barreiro Javier, Didangelos Athanasios, Yin Xiaoke, Doménech Nieves, Mayr Manuel

机构信息

King's British Heart Foundation Centre, London, UK.

出版信息

Methods Mol Biol. 2013;1005:215-23. doi: 10.1007/978-1-62703-386-2_17.

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.

摘要

心脏纤维化的特征是细胞外基质(ECM)过度沉积,是各种心血管疾病的常见并发症。然而,人们对心脏细胞外空间中的蛋白质了解甚少。由于存在更丰富的细胞内蛋白质、完整ECM蛋白质的低溶解度以及大量的翻译后修饰,对心脏ECM进行蛋白质组学分析可能具有挑战性。在这里,我们描述了一种基于组织去细胞化的提取方法,该方法允许对心脏组织中的细胞外蛋白质进行生化亚分级分离。这些相对低复杂性的蛋白质级分适用于通过凝胶液相色谱-串联质谱(gel-LC-MS/MS)和其他蛋白质组学技术进行分析。

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