Large-Scale Characterization and Analysis of the Murine Cardiac Proteome
Large-Scale Characterization and Analysis of the Murine Cardiac Proteome
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DOI：
10.1021/pr800845a
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发表时间：
2009-04-01
影响因子：
4.4
通讯作者：
Gramolini, Anthony O.
中科院分区：
文献类型：
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作者：
Bousette, Nicolas;Kislinger, Thomas;Gramolini, Anthony O.
质谱和生物信息学的最新进展提供了表征来自各种生物体和细胞类型的复杂蛋白质景观的手段。标准蛋白质组的发展，展示了所有的蛋白质参与正常生理将有助于疾病机制的描绘。在这里，我们研究了野生型心脏蛋白质组使用的数据从亚细胞分馏协议结合多维蛋白质鉴定蛋白质组学方法。我们确定了4906蛋白质被分配到细胞溶质，微粒体，线粒体基质或线粒体膜组分在每个馏分中的相对丰度值。我们对这些蛋白质进行了层次聚类，基因本体论术语分析，免疫印迹，与公开可用的蛋白质数据库进行比较，与4种不同的心脏转录组进行比较，最后与6种其他相关的蛋白质组数据集进行比较。这项研究提供了一个详尽的分析心脏蛋白质组，是第一次大规模调查的亚细胞位置超过2000未注释的蛋白质。随着消减转录组学方法的使用，我们还扩展了我们的分析，以确定我们的蛋白质组中的“心脏选择性”因子。最后，使用特定的过滤标准，我们确定了蛋白质型肽，随后用于小鼠和人的靶向研究。因此，我们提供这作为一个重大贡献，在心血管研究的蛋白质组学领域的进步。
Recent advances in mass spectrometry and bioinformatics have provided the means to characterize complex protein landscapes from a wide variety of organisms and cell types. Development of standard proteomes exhibiting all of the proteins involved in normal physiology will facilitate the delineation of disease mechanisms. Here, we examine the wild-type cardiac proteome using data obtained from a subcellular fractionation protocol in combination with a multidimensional protein identification proteomics approach. We identified 4906 proteins which were allocated to either cytosolic, microsomal, mitochondrial matrix or mitochondrial membrane fractions with relative abundance values in each fraction. We subjected these proteins to hierarchical clustering, gene ontology terms analysis, immunoblotting, comparison to publicly available protein databases, comparison to 4 distinct cardiac transcriptomes, and finally, to 6 other related proteomic data sets. This study provides an exhaustive analysis of the cardiac proteome and is the first large-scale investigation of the subcellular location for over 2000 unannotated proteins. With the use of a subtractive transcriptomics approach, we have also extended our analysis to identify 'cardiac selective' factors in our proteome. Finally, using specific filtering criteria, we identified proteotypic peptides for subsequent use in targeted studies of both mouse and human. Therefore, we offer this as a major contribution to the advancement of the field of proteomics in cardiovascular research.