Quantitative profile of five murine core proteomes using label-free functional proteomics
Quantitative profile of five murine core proteomes using label-free functional proteomics
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DOI：
10.1074/mcp.m700037-mcp200
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发表时间：
2007-09-01
影响因子：
7
通讯作者：
Vanhaesebroeck, Bart
中科院分区：
文献类型：
--
作者：
Cutillas, Pedro R.;Vanhaesebroeck, Bart
对动物和人体组织的分析是生物学和生物医学研究的关键。初级生物材料的比较蛋白质组学分析将受益于能够评估无限数量样品的简单实验工作流程。在这份报告中，我们描述了应用无标记蛋白质组学的定量分析的五个小鼠核心蛋白质组。我们开发了一个计算机程序和标准化程序，允许利用LC-MS/ MS实验中固有的定量数据，用于复杂混合物中蛋白质的相对和绝对定量。该方法的重要特征包括（i）其比较无限数量的样品的能力，（ii）其对原代组织和培养细胞的适用性，（iii）其简单的工作流程而无需化学反应步骤，以及（iv）其不仅用于相对定量而且用于估计绝对蛋白质丰度的有用性。我们应用这种方法来定量表征小鼠脑，心脏，肾脏，肝脏和肺中最丰富的蛋白质。我们将8，800个MS/ MS肽谱与1，500个蛋白质进行了匹配，并生成了44，000个独立的数据点，以分析小鼠组织中1，000种最丰富的蛋白质。该数据集提供了小鼠基本蛋白质组的定量图谱，鉴定了器官特异性蛋白质组之间的主要相似性和差异，并作为无标记定量MS如何用于在分子水平上表征哺乳动物原代组织表型的范例。
Analysis of primary animal and human tissues is key in biological and biomedical research. Comparative proteomics analysis of primary biological material would benefit from uncomplicated experimental work flows capable of evaluating an unlimited number of samples. In this report we describe the application of label- free proteomics to the quantitative analysis of five mouse core proteomes. We developed a computer program and normalization procedures that allow exploitation of the quantitative data inherent in LC- MS/ MS experiments for relative and absolute quantification of proteins in complex mixtures. Important features of this approach include ( i) its ability to compare an unlimited number of samples, ( ii) its applicability to primary tissues and cultured cells, ( iii) its straightforward work flow without chemical reaction steps, and ( iv) its usefulness not only for relative quantification but also for estimation of absolute protein abundance. We applied this approach to quantitatively characterize the most abundant proteins in murine brain, heart, kidney, liver, and lung. We matched 8,800 MS/ MS peptide spectra to 1,500 proteins and generated 44,000 independent data points to profile the similar to 1,000 most abundant proteins in mouse tissues. This dataset provides a quantitative profile of the fundamental proteome of a mouse, identifies the major similarities and differences between organ- specific proteomes, and serves as a paradigm of how label- free quantitative MS can be used to characterize the phenotype of mammalian primary tissues at the molecular level.