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  A Mass Spectrometry-Based Approach for Mapping Protein Subcellular Localization Reveals the Spatial Proteome of Mouse Primary Neurons

Itzhak, D. N., Davies, C., Tyanova, S., Mishra, A., Williamson, J., Antrobus, R., et al. (2017). A Mass Spectrometry-Based Approach for Mapping Protein Subcellular Localization Reveals the Spatial Proteome of Mouse Primary Neurons. Cell Reports, 20(11), 2706-2718. doi:10.1016/j.celrep.2017.08.063.

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 Creators:
Itzhak, Daniel N.1, Author              
Davies, Colin2, Author
Tyanova, Stefka1, Author              
Mishra, Archana3, Author              
Williamson, James2, Author
Antrobus, Robin2, Author
Cox, Jürgen1, Author              
Weekes, Michael P.2, Author
Borner, Georg H. H.1, Author              
Affiliations:
1Mann, Matthias / Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565159              
2external, ou_persistent22              
3Department: Molecules-Signaling-Development / Klein, MPI of Neurobiology, Max Planck Society, ou_1113546              

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Free keywords: COMPUTATIONAL PLATFORM; ACCURATE; MAP; QUANTIFICATION; EXPRESSION; CELLSCell Biology;
 Abstract: 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.

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Language(s): eng - English
 Dates: 2017
 Publication Status: Published in print
 Pages: 13
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Degree: -

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Title: Cell Reports
Source Genre: Journal
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Publ. Info: Maryland Heights, MO : Cell Press
Pages: - Volume / Issue: 20 (11) Sequence Number: - Start / End Page: 2706 - 2718 Identifier: Other: 2211-1247
CoNE: https://pure.mpg.de/cone/journals/resource/2211-1247