A Mass Spectrometry-Based Approach for Mapping Protein Subcellular Localization Reveals the Spatial Proteome of Mouse Primary Neurons.
A Mass Spectrometry-Based Approach for Mapping Protein Subcellular Localization Reveals the Spatial Proteome of Mouse Primary Neurons.
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基于质谱的方法用于绘制蛋白质亚细胞定位，揭示了小鼠原代神经元的空间蛋白质组。
DOI：
10.1016/j.celrep.2017.08.063
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发表时间：
2017-09-12
期刊：
影响因子：
8.8
通讯作者：
Borner GHH
中科院分区：
文献类型：
--
作者：
Itzhak DN;Davies C;Tyanova S;Mishra A;Williamson J;Antrobus R;Cox J;Weekes MP;Borner GHH
我们以前开发了一种基于质谱学的方法，动态细胞器图谱，用于在比较实验中确定蛋白质亚细胞定位和识别易位事件。代谢标记用于定量(细胞培养中氨基酸的稳定同位素标记[SILAC])的使用使该方法最适合于在培养中生长的细胞。在这里，我们调整了工作流程，以适应无标记量化(LFQ)和化学标记/多重策略(串联质量标记[TMT])。这两种方法对于细胞器图谱的生成和蛋白质易位的捕获都是非常有效的。此外，将无标记细胞器映射应用于急性分离的小鼠初级神经元，提供了超过8,000种蛋白质的亚细胞定位和拷贝数信息，使详细分析细胞器组织成为可能。我们的研究将动态细胞器图谱的范围扩展到任何细胞类型或组织，也扩展到高通量筛选。高分辨率细胞器图谱与无标记量化(LFQ)高通量细胞质图谱基于TMT的多路复用EGF诱导的蛋白质定位变化与SILAC、LFQ和TMT的深度定位来自小鼠原代神经元的定量空间蛋白质组动态细胞器图谱先前为捕捉培养细胞中蛋白质亚细胞定位变化提供了蛋白质组学方法。Itzhak等人。现在已将该方法调整为通用格式，将该方法扩展到所有类型的单元格。对原代小鼠神经元的应用提供了超过8,000种蛋白质的空间和定量信息。
We previously developed a mass spectrometry-based method, dynamic organellar maps, for the determination of protein subcellular localization and identification of translocation events in comparative experiments. The use of metabolic labeling for quantification (stable isotope labeling by amino acids in cell culture [SILAC]) renders the method best suited to cells grown in culture. Here, we have adapted the workflow to both label-free quantification (LFQ) and chemical labeling/multiplexing strategies (tandem mass tagging [TMT]). Both methods are highly effective for the generation of organellar maps and capture of protein translocations. Furthermore, application of label-free organellar mapping to acutely isolated mouse primary neurons provided subcellular localization and copy-number information for over 8,000 proteins, allowing a detailed analysis of organellar organization. Our study extends the scope of dynamic organellar maps to any cell type or tissue and also to high-throughput screening. High-resolution organellar maps with label-free quantification (LFQ) High-throughput organellar maps with TMT-based multiplexing Deep mapping of EGF-induced protein localization changes with SILAC, LFQ, and TMT A quantitative spatial proteome from mouse primary neurons Dynamic organellar maps previously provided a proteomic method for capturing protein subcellular localization changes in cultured cells. Itzhak et al. have now adapted the approach to a universal format, extending the method to all cell types. Application to primary mouse neurons provides spatial and quantitative information for more than 8,000 proteins.
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影响因子：
--
作者：
Mardakheh FK;Sailem HZ;Kümper S;Tape CJ;McCully RR;Paul A;Anjomani-Virmouni S;Jørgensen C;Poulogiannis G;Marshall CJ;Bakal C
通讯作者：
Bakal C
影响因子：
16.6
作者：
Christoforou A;Mulvey CM;Breckels LM;Geladaki A;Hurrell T;Hayward PC;Naake T;Gatto L;Viner R;Martinez Arias A;Lilley KS
通讯作者：
Lilley KS
DOI：
10.1007/978-1-4939-6747-6_10
发表时间：
2017-01-01
期刊：
PROTEOMICS: METHODS AND PROTOCOLS
影响因子：
--
作者：
Hesketh, Geoffrey G.;Youn, Ji-Young;Gingras, Anne-Claude
通讯作者：
Gingras, Anne-Claude
影响因子：
64.5
作者：
Foster, LJ;de Hoog, CL;Mann, M
通讯作者：
Mann, M
影响因子：
7
作者：
Jadot, Michel;Boonen, Marielle;Lobel, Peter
通讯作者：
Lobel, Peter