High Quality Catalog of Proteotypic Peptides from Human Heart

被引:34
作者
Kline, Kelli G. [2 ]
Frewen, Barbara [1 ]
Bristow, Michael R. [3 ]
MacCoss, Michael J. [1 ]
Wu, Christine C. [2 ]
机构
[1] Univ Washington, Dept Genome Sci, Seattle, WA 98101 USA
[2] Univ Colorado, Sch Med, Dept Pharmacol, Aurora, CO 80045 USA
[3] Univ Colorado, Sch Med, Dept Cardiol, Aurora, CO 80045 USA
基金
美国国家卫生研究院;
关键词
proteotypic peptides; targeted mass spectrometry; human heart explant; dilated cardiomyopathy; MudPIT;
D O I
10.1021/pr800239e
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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 nonfailing and failing human heart explants were optimized for shotgun proteomic analysis using Multidimensional Protein Identification Technology (MudPIT). Ten replicate MudPIT analyses were performed on each tissue sample and resulted in the identification of 30 605 unique peptides with a q-value <= 0.01, corresponding to 7138 unique human heart proteins. Experimental observation frequencies were assessed and used to select over 4476 proteotypic peptides for 2558 heart proteins. This human cardiac data set 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.
引用
收藏
页码:5055 / 5061
页数:7
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