Large-scale analysis of the human ubiquitin-related proteome
Large-scale analysis of the human ubiquitin-related proteome
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DOI：
10.1002/pmic.200401280
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发表时间：
2005-11-01
期刊：
影响因子：
3.4
通讯作者：
Nakayama, KI
中科院分区：
文献类型：
--
作者：
Matsumoto, M;Hatakeyama, S;Nakayama, KI
蛋白质泛素化有助于调节许多细胞过程，包括蛋白质降解、受体内化和DNA损伤修复。我们现在提出了一个在人类细胞中泛素结合蛋白和泛素相关蛋白的全面特征。在变性和自然条件下，用免疫亲和层析对蛋白进行纯化。其中345个(51%)蛋白质被归类为URP-D(变性条件下泛素相关蛋白质组)，325个(49%)蛋白质被归类为URP-N(仅在自然条件下与泛素相关的蛋白质组)，其中325个蛋白质(49%)包括泛素化蛋白质相关分子。URP-D和URP-N在不同功能类别中的蛋白质比例有很大差异。在URP-D组蛋白中检测到许多核糖体亚基，质谱分析直接证明其中几个亚基是泛素化的，这表明泛素化可能在核糖体蛋白的调节和/或质量控制中发挥重要作用。我们的结果证明了蛋白质泛素化的蛋白质组学分析的潜力，为调节蛋白质稳定性和其他泛素相关的细胞功能提供了重要的见解。
Protein ubiquitylation contributes to the regulation of many cellular processes including protein degradation, receptor internalization, and repair of DNA damage. We now present a comprehensive characterization of ubiquitin-conjugated and ubiquitin-associated proteins in human cells. The proteins were purified by immunoaffinity chromatography under denaturing or native conditions. They were then digested with trypsin, and the resulting peptides were analyzed by 2-D LC and MS/MS. A total of 670 distinct proteins were identified; 345 proteins (51%) were classified as Urp-D (ubiquitin-related proteome under the denaturing condition) and comprised ubiquitin-conjugated molecules, whereas 325 proteins (49%) were classified as Urp-N (ubiquitin-related proteome only under the native condition) and included molecules that associated with ubiquitylated proteins. The proportions of proteins in various functional categories differed substantially between Urp-D and Urp-N. Many ribosomal subunits were detected in the Urp-D group of proteins and several of these subunits were directly shown to be ubiquitylated by mass spectrometric analysis, suggesting that ubiquitylation might play an important role in the regulation and/or quality control of ribosomal proteins. Our results demonstrate the potential of proteomics analysis of protein ubiquitylation to provide important insight into the regulation of protein stability and other ubiquitin-related cellular functions.