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Activity-Based Protein Profiling with Natural Product-Derived Chemical Probes in Human Cell Lysates

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Activity-Based Proteomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1491))

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Abstract

Bioreactive natural products represent versatile starting points for the development of structurally unique activity-based probes. In the present protocol, we describe the workflow for an activity-based protein profiling (ABPP) experiment with an alkyne-tagged natural product derivative. Our protocol includes experimental procedures for in vivo labeling, sample preparation and 2-step (click chemistry) visualization and sample preparation for mass spectrometry-based target identification.

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Acknowledgements

The authors would like to acknowledge financial support from the following funding agencies: ERC starting grant (to M.K., grant No. 258413), the COST Action CM1004 and the DFG (to M.K., grants no. INST 20876/127-1 FUGG and SFB 1093/1).

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Authors and Affiliations

  1. ZMB, Faculty of Biology, University of Duisburg-Essen, Universitätsstr. 2, 45117, Essen, Germany

    Susanne Zweerink, Tanja Pollmann, Sabrina Ninck, Farnusch Kaschani & Markus Kaiser

Authors
  1. Susanne Zweerink
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  2. Tanja Pollmann
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  3. Sabrina Ninck
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  4. Farnusch Kaschani
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  5. Markus Kaiser
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Corresponding author

Correspondence to Markus Kaiser .

Editor information

Editors and Affiliations

  1. Bio-Organic Synthesis Group Leiden Institute of Chemistry, Leiden University, Leiden, The Netherlands

    Herman S. Overkleeft

  2. Bio-Organic Synthesis Group Leiden Institute of Chemistry, Leiden University, Leiden, The Netherlands

    Bogdan I. Florea

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Zweerink, S., Pollmann, T., Ninck, S., Kaschani, F., Kaiser, M. (2017). Activity-Based Protein Profiling with Natural Product-Derived Chemical Probes in Human Cell Lysates. In: Overkleeft, H., Florea, B. (eds) Activity-Based Proteomics. Methods in Molecular Biology, vol 1491. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6439-0_3

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  • DOI: https://doi.org/10.1007/978-1-4939-6439-0_3

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6437-6

  • Online ISBN: 978-1-4939-6439-0

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