HIGH QUALITY CATALOG OF PROTEOTYPIC PEPTIDES FROM HUMAN HEART

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 蛋白质组学研究开始扩展到更传统的蛋白质混合物的鸟枪分析，包括使用质谱法对特定蛋白质的靶向分析。 基于靶向质谱法的稳健测定法的开发不可或缺的是哪些肽提供了原始基因产物（即蛋白质型肽）的准确和灵敏的替代物的先验知识。 开发一个目录的蛋白质型肽在人类心脏，TRIzol？使用多维蛋白质鉴定技术（MudPIT）优化来自非衰竭和衰竭的人心脏外植体的左心室组织提取物用于鸟枪蛋白质组学分析。 对每个组织样品进行10次重复MudPIT分析，结果鉴定出30，605种独特的肽，q值为0.01或更好，对应于7，138种独特的人类心脏蛋白。 评估实验观察频率，并用于为2，558种心脏蛋白质选择超过4，476种蛋白质型肽。 这个人类心脏数据集可以作为一个公共参考，以指导未来的靶向质谱实验监测人类心脏疾病的潜在蛋白质生物标志物的蛋白质型肽的选择。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Proteomics research is beginning to expand beyond the more traditional shotgun analysis of protein mixtures to include targeted analyses of specific proteins using mass spectrometry. Integral to the development of a robust assay based on targeted mass spectrometry is prior knowledge of which peptides provide an accurate and sensitive proxy of the originating gene product (i.e. proteotypic peptides). To develop a catalog of proteotypic peptides in human heart, TRIzol? extracts of left-ventricular tissue from non-failing and failing human heart explants were optimized for shotgun proteomic analysis using Multi-dimensional Protein Identification Technology (MudPIT). Ten replicate MudPIT analyses were performed on each tissue sample and resulted in the identification 30,605 unique peptides with a q-value of 0.01 or better, corresponding to 7,138 unique human heart proteins. Experimental observation frequencies were assessed and used to select over 4,476 proteotypic peptides for 2,558 heart proteins. This human cardiac dataset can serve as a public reference to guide the selection of proteotypic peptides for future targeted mass spectrometry experiments monitoring potential protein biomarkers of human heart diseases.