Global subcellular characterization of protein degradation using quantitative proteomics.
Global subcellular characterization of protein degradation using quantitative proteomics.
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DOI：
10.1074/mcp.m112.024547
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发表时间：
2013-03
期刊：
影响因子：
--
通讯作者：
Lamond AI
中科院分区：
文献类型：
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作者：
Larance M;Ahmad Y;Kirkwood KJ;Ly T;Lamond AI
蛋白质降解提供了用于控制细胞周期进程和许多其他细胞途径的重要调节机制。为了全面分析 U2OS 骨肉瘤细胞中蛋白质降解的空间控制，我们将药物治疗和基于 SILAC 的定量质谱与亚细胞和蛋白质分级相结合。由此产生的数据集分析了来自核、细胞质、膜和细胞骨架区室的超过 74,000 种肽，对应于 ∼5000 种蛋白质。这些数据确定了快速降解的蛋白酶体靶标，例如 PRR11，并强调了通过阻断蛋白酶体诱导的导致翻译抑制的反馈机制。我们证明这是由未折叠蛋白反应的激活介导的。我们观察到蛋白质降解的区室特异性差异，包括通过全细胞裂解物分析不会被快速降解的蛋白质。整个数据集的生物信息分析显示在蛋白质组动力学百科全书中，这是一个基于网络的资源，其中蛋白质注释了稳定性和亚细胞分布。
Protein degradation provides an important regulatory mechanism used to control cell cycle progression and many other cellular pathways. To comprehensively analyze the spatial control of protein degradation in U2OS osteosarcoma cells, we have combined drug treatment and SILAC-based quantitative mass spectrometry with subcellular and protein fractionation. The resulting data set analyzed more than 74,000 peptides, corresponding to ∼5000 proteins, from nuclear, cytosolic, membrane, and cytoskeletal compartments. These data identified rapidly degraded proteasome targets, such as PRR11 and highlighted a feedback mechanism resulting in translation inhibition, induced by blocking the proteasome. We show this is mediated by activation of the unfolded protein response. We observed compartment-specific differences in protein degradation, including proteins that would not have been characterized as rapidly degraded through analysis of whole cell lysates. Bioinformatic analysis of the entire data set is presented in the Encyclopedia of Proteome Dynamics, a web-based resource, with proteins annotated for stability and subcellular distribution.