Micro-proteomics with iterative data analysis: Proteome analysis in C. elegans at the single worm level.
Micro-proteomics with iterative data analysis: Proteome analysis in C. elegans at the single worm level.
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DOI：
10.1002/pmic.201500264
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发表时间：
2016-02
期刊：
影响因子：
3.4
通讯作者：
Lamond AI
中科院分区：
文献类型：
--
作者：
Bensaddek D;Narayan V;Nicolas A;Murillo AB;Gartner A;Kenyon CJ;Lamond AI
蛋白质组学研究通常在群体水平上分析蛋白质，使用从数万到数百万个细胞中提取的提取物。由此产生的测量值对应于整个细胞群的平均值，并且可以掩盖单个细胞或生物体之间蛋白质表达和功能的相当大的差异。在这里，我们报告了用于秀丽隐杆线虫（一种由959个体细胞和约1500个生殖细胞组成的真核生物）分析的微蛋白质组学的发展，在单个生物体水平上测量蠕虫蛋白质组，深度为约3000个蛋白质。这包括在广泛的动态表达水平范围内（bbb6数量级）检测蛋白质，包括许多参与染色体结构和基因调控的染色质相关因子。我们应用微蛋白质组学工作流程来测量单个线虫对热休克的整体蛋白质组反应。这显示了个体动物在热休克后蛋白质组反应程度的差异，包括热休克蛋白诱导的不同。因此，微蛋白质组学管道有助于研究秀丽隐杆线虫等基因群体中个体之间蛋白质表达的随机变化。本研究中描述的所有数据都可以通过蛋白质组动力学百科全书（http://www.peptracker.com/epd）在线获得，这是一个开放获取，可搜索的数据库资源。
Proteomics studies typically analyze proteins at a population level, using extracts prepared from tens of thousands to millions of cells. The resulting measurements correspond to average values across the cell population and can mask considerable variation in protein expression and function between individual cells or organisms. Here, we report the development of micro‐proteomics for the analysis of Caenorhabditis elegans, a eukaryote composed of 959 somatic cells and ∼1500 germ cells, measuring the worm proteome at a single organism level to a depth of ∼3000 proteins. This includes detection of proteins across a wide dynamic range of expression levels (>6 orders of magnitude), including many chromatin‐associated factors involved in chromosome structure and gene regulation. We apply the micro‐proteomics workflow to measure the global proteome response to heat‐shock in individual nematodes. This shows variation between individual animals in the magnitude of proteome response following heat‐shock, including variable induction of heat‐shock proteins. The micro‐proteomics pipeline thus facilitates the investigation of stochastic variation in protein expression between individuals within an isogenic population of C. elegans. All data described in this study are available online via the Encyclopedia of Proteome Dynamics (http://www.peptracker.com/epd), an open access, searchable database resource.