Direct Proteomic Quantification of the Secretome of Activated Immune Cells
Direct Proteomic Quantification of the Secretome of Activated Immune Cells
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DOI：
10.1126/science.1232578
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发表时间：
2013-04-26
期刊：
影响因子：
56.9
通讯作者：
Mann, Matthias
中科院分区：
文献类型：
--
作者：
Meissner, Felix;Scheltema, Richard A.;Mann, Matthias
蛋白质的分泌允许生物体中远处细胞的交流，并控制着广泛的生理功能。我们描述了一个定量的，高分辨率的质谱工作流程，以检测和量化受体连接后从免疫细胞释放的蛋白质。我们量化了775种蛋白的时间分辨释放，包括52种带注释的细胞因子，这些细胞来自150000个初级toll样受体4激活的巨噬细胞。实现低picogram sensitivity，我们检测到的分泌蛋白的丰度在刺激下增加了1万多倍。分泌组与转录组的比较揭示了溶酶体蛋白的转录解耦释放。从遗传模型中，我们定义了依赖于不同细胞内信号接头的分泌谱，并表明许多促炎蛋白的分泌受到冗余机制的保护，而信号接头的协同作用促进了抗炎蛋白的释放。
Protein secretion allows communication of distant cells in an organism and controls a broad range of physiological functions. We describe a quantitative, high-resolution mass spectrometric workflow to detect and quantify proteins that are released from immune cells upon receptor ligation. We quantified the time-resolved release of 775 proteins, including 52 annotated cytokines from only 150,000 primary Toll-like receptor 4-activated macrophages per condition. Achieving low picogram sensitivity, we detected secreted proteins whose abundance increased by a factor of more than 10,000 upon stimulation. Secretome to transcriptome comparisons revealed the transcriptionally decoupled release of lysosomal proteins. From genetic models, we defined secretory profiles that depended on distinct intracellular signaling adaptors and showed that secretion of many proinflammatory proteins is safeguarded by redundant mechanisms, whereas signaling adaptor synergy promoted the release of anti-inflammatory proteins.