Yabluchanskiy Andriy, Li Yaojun, de Castro Brás Lisandra E, Hakala Kevin, Weintraub Susan T, Lindsey Merry L
San Antonio Cardiovascular Proteomics Center, The University of Texas Health Science Center, San Antonio, TX, USA.
Methods Mol Biol. 2013;1066:185-99. doi: 10.1007/978-1-62703-604-7_16.
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.
心肌梗死后左心室重构涉及多种调节修复反应的机制。基质金属蛋白酶（MMPs）是协调细胞外基质周转的主要蛋白水解酶家族。MMP-7或MMP-9缺失可减轻心肌梗死后的不良重构，但其机制尚未完全阐明。MMP-7和MMP-9都有大量已知的体外底物，但在心肌梗死情况下这两种MMP的体内底物尚未完全确定。蛋白质组学技术的进步使得对细胞和组织中的蛋白质表达进行全面分析成为可能。在本章中，我们描述了一种使用二维电泳、液相色谱-串联质谱和免疫印迹对心肌梗死后左心室中体内候选MMP底物进行蛋白质组学分析的方案。
