HIGH QUALITY CATALOG OF PROTEOTYPIC PEPTIDES FROM HUMAN HEART

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 蛋白质组学研究正开始超越对蛋白质混合物的更传统的鸟枪式分析，包括使用质谱学对特定蛋白质进行有针对性的分析。建立基于靶向质谱学的可靠分析方法的关键是事先知道哪些多肽能够准确和灵敏地替代原始基因产物(即蛋白质型多肽)。为了开发一个人类心脏蛋白类型肽的目录，Trizol？利用多维蛋白质鉴定技术(MudPIT)对非衰竭和衰竭心脏组织的提取液进行了优化，以用于鸟枪式蛋白质组学分析。对每个组织样本进行了10次重复的MudPIT分析，结果鉴定出30,605个Q值为0.01或更高的独特多肽，对应于7,138个独特的人类心脏蛋白。实验观察频率被评估并用于为2,558个心脏蛋白选择超过4,476个蛋白类型肽。这一人类心脏数据集可以作为公共参考，指导为未来的靶向质谱学实验选择蛋白质型肽，以监测人类心脏病的潜在蛋白质生物标志物。

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.