Antibody-based protein profiling of the human chromosome 21.
Antibody-based protein profiling of the human chromosome 21.
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DOI：
10.1074/mcp.m111.013458
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发表时间：
2012-03
期刊：
影响因子：
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通讯作者：
Sivertsson Å
中科院分区：
文献类型：
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作者：
Uhlén M;Oksvold P;Älgenäs C;Hamsten C;Fagerberg L;Klevebring D;Lundberg E;Odeberg J;Pontén F;Kondo T;Sivertsson Å
已经提出了人类蛋白质组项目，以以基因为中心，在所有人类蛋白质的系统图谱的基础上，逐条染色体地创建一个基于知识的资源。在此背景下，我们在这里描述了对21号染色体的系统分析，使用基于抗体的方法进行蛋白质谱分析，使用共聚焦显微镜和免疫组织化学，并辅以使用下一代测序数据的转录本谱分析。我们还描述了一种结合基于抗体的探测和等电聚焦来分析蛋白质异构体的新方法。这项分析已经在21号染色体上发现了几个基因，而此前在蛋白质水平上没有证据，而亚型分析表明，很大一部分人类蛋白质具有多种亚型。关于相应蛋白质的亚细胞定位、组织分布和分子特征，给出了染色体范围内所有21号染色体基因的状态。讨论了产生染色体特定资源的途径，包括来自互补分析平台的集成数据，例如质谱学和基因标签分析。
The Human Proteome Project has been proposed to create a knowledge-based resource based on a systematical mapping of all human proteins, chromosome by chromosome, in a gene-centric manner. With this background, we here describe the systematic analysis of chromosome 21 using an antibody-based approach for protein profiling using both confocal microscopy and immunohistochemistry, complemented with transcript profiling using next generation sequencing data. We also describe a new approach for protein isoform analysis using a combination of antibody-based probing and isoelectric focusing. The analysis has identified several genes on chromosome 21 with no previous evidence on the protein level, and the isoform analysis indicates that a large fraction of human proteins have multiple isoforms. A chromosome-wide matrix is presented with status for all chromosome 21 genes regarding subcellular localization, tissue distribution, and molecular characterization of the corresponding proteins. The path to generate a chromosome-specific resource, including integrated data from complementary assay platforms, such as mass spectrometry and gene tagging analysis, is discussed.