基于等压标记的液相色谱-串联质谱策略用于人胰腺组织蛋白质组的全面分析
Isobaric Labeling-Based LC-MS/MS Strategy for Comprehensive Profiling of Human Pancreatic Tissue Proteome.
作者信息
Liu Chih-Wei, Zhang Qibin
机构信息
Center for Translational Biomedical Research, University of North Carolina at Greensboro, North Carolina Research Campus, 500 Laureate Way Suite 4226, Kannapolis, NC, 28081, USA.
Department of Chemistry and Biochemistry, University of North Carolina at Greensboro, Greensboro, NC, USA.
出版信息
Methods Mol Biol. 2018;1788:215-224. doi: 10.1007/7651_2017_77.
The pancreas is an organ with both endocrine and exocrine functions, and various pathologies, such as pancreatic cancer and diabetes are associated with this organ. Owing to the limited pancreatic biopsy samples available for research, it is critical to make the best use of cadaveric pancreatic tissue for biomarker studies and mechanistic understanding of pancreas-related pathologies. Discovery-phase quantitative proteomics has attracted a lot of attention for its capabilities in large-scale protein identification and accurate protein quantification. Here, we describe a workflow using isobaric labeling (tandem mass tag or TMT) based quantitative proteomics to confidently identify and quantify human pancreatic tissue proteome, including sample preparation, isobaric tag labeling, peptide level fractionation, LC-MS/MS, database search, and statistical analysis.
胰腺是一个兼具内分泌和外分泌功能的器官，胰腺癌和糖尿病等各种病症都与该器官有关。由于可用于研究的胰腺活检样本有限，充分利用尸体胰腺组织进行生物标志物研究以及对胰腺相关病症的机制理解至关重要。发现阶段的定量蛋白质组学因其在大规模蛋白质鉴定和准确蛋白质定量方面的能力而备受关注。在此，我们描述了一种基于等压标记（串联质谱标签或TMT）的定量蛋白质组学工作流程，用于可靠地鉴定和定量人胰腺组织蛋白质组，包括样品制备、等压标签标记、肽段水平分级分离、液相色谱-串联质谱、数据库搜索和统计分析。
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