Proteome-wide protein concentrations in the human heart
Proteome-wide protein concentrations in the human heart
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DOI：
10.1039/c004495d
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发表时间：
2010-01-01
影响因子：
--
通讯作者：
Heck, Albert J. R.
中科院分区：
文献类型：
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作者：
Aye, Thin Thin;Scholten, Arjen;Heck, Albert J. R.
人体心脏的最大组成部分，左心室（LV），在整个身体的血液输送中起着重要作用。因此，深入详细的定量蛋白质组分析的人左心室是一个宝贵的资源。为此，使用了一种多方面的蛋白质组学方法，该方法结合了差异样品分级（凝胶、强阳离子交换（SCX））、酶消化（胰蛋白酶、胰凝乳蛋白酶、LysN）和肽片段化技术（CID和ETcaD），以提高蛋白质序列覆盖率、鉴定置信度和定量丰度测定。使用严格的标准，3584个不同的蛋白质可以从最新的良好注释的Swissprot数据库（23 000个条目）。此外，使用超过130 000个已鉴别的MS/MS光谱，通过光谱计数方法的组合评估每种已鉴别的LV蛋白的浓度。在最集中的蛋白质中，存在许多目前用于检测心肌梗死的生物标志物。这些心漏标志物具有良好的诊断能力，但其预后潜力似乎有限。发现代表心脏病病因决定因素的标志物需要将焦点转移到信号蛋白质组。因此，采用以蛋白质类为中心的激酶、磷酸酶和GTP酶定量分析。这些比较分析揭示了许多心脏相关的激酶（PKA，CaMKII，ERK）驻留在最丰富的信号蛋白，也介导许多观察到的体内磷酸化位点。信号蛋白的丰度图可能有助于识别新的功能途径，例如通过丰富但相对鲜为人知的激酶STK 38 L和OXSR 1。所获得的人左心室的定量蛋白质文库是分离基于信号传导的推定生物标志物的有价值的资源，其浓度可能在血浆中可检测到。
The largest component of the human heart, the left ventricle (LV), plays a major role in delivering blood throughout the body. Therefore, an in-depth detailed quantitative proteome analysis of the human LV is a valuable resource. For this purpose, a multifaceted proteomics approach combining differential sample fractionations (gel, strong cation exchange (SCX)), enzymatic digestions (trypsin, chymotrypsin, LysN), and peptide fragmentation techniques (CID and ETcaD) was used to enhance protein sequence coverage, identification confidence and quantitative abundance determination. Using stringent criteria, 3584 distinct proteins could be identified from the latest well-annotated Swissprot database (23 000 entries). Commutatively, the over 130 000 identified MS/MS spectra were used to assess concentrations of each identified LV protein through a combination of spectral counting methods. Among the most concentrated proteins, many currently used biomarkers for detection of myocardial infarction reside. These cardiac leakage markers have a good diagnostic power, but their prognostic potential seems limited. Discovery of markers that represent etiological determinants of cardiac disease require a shift of focus towards the signaling proteome. Therefore, a protein-class centered quantitative analysis of kinases, phosphatases and GTPases was adopted. These comparative analyses revealed many cardiac involved kinases (PKA, CaMKII, ERK) to reside among the most abundant signaling proteins, and also to mediate many observed in vivo phosphorylation sites. The abundance chart of signaling proteins may assist in identifying novel functional pathways, for instance through the abundant, but relatively little known, kinases STK38L and OXSR1. The obtained quantitative protein library of the human left ventricle is a valuable resource to isolate signaling based, putative biomarkers with concentrations likely to be detectable in plasma.