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Post-Translational Modification Profiling-Functional Proteomics for the Analysis of Immune Regulation

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Proteomics for Drug Discovery

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1647))

Abstract

Posttranslational modifications (PTMs) of proteins are an integral part of major cellular regulatory mechanisms dictating protein function, localization, and stability. The capacity to screen PTMs using protein microarrays has advanced our ability to identify their targets and regulatory role. This chapter discusses a unique procedure that combines functional extract-based activity assay with large-scale screening utilities of protein microarrays. This “PTM-profiling” system offers advantages in quantitatively identifying modifications in an unbiased manner in the context of specific cellular conditions. While the possibilities of studying PTMs in different settings are enormous, the immune system presents an attractive model for studying the effects of perturbations in PTMs, and specifically the ubiquitin system, as these were already implicated in both immune function and dysfunction. This chapter discusses the significance of PTM profiling in addressing basic questions in immunology. We describe detailed protocols for the preparation of functional cell extracts from immune cell cultures, following differentiation or induced signals, and screening PTMs on protein arrays, as well as basic guidelines for data analysis and interpretation.

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Acknowledgments

Yifat Merbl is supported by the I-CORE Program of the Planning and Budgeting Committee and The Israel Science Foundation (grant No. 1775/12), and by the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme.

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Authors and Affiliations

  1. Department of Immunology, The Weizmann Institute of Science, 234 Herzl Street, Rehovot, 7610001, Israel

    Avital Eisenberg-Lerner, Ifat Regev & Yifat Merbl

Authors
  1. Avital Eisenberg-Lerner
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  2. Ifat Regev
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  3. Yifat Merbl
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Corresponding authors

Correspondence to Avital Eisenberg-Lerner or Yifat Merbl .

Editor information

Editors and Affiliations

  1. Department of Biological Sciences, Virginia Tech, Blacksburg, Virginia, USA

    Iulia M. Lazar

  2. Department of Pharmaceutical Sciences, School of Pharmacy, Southern Illinois University Edwardsville, Edwardsville, Illinois, USA

    Maria Kontoyianni

  3. ImmunoGen, Waltham, Massachusetts, USA

    Alexandru C. Lazar

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Eisenberg-Lerner, A., Regev, I., Merbl, Y. (2017). Post-Translational Modification Profiling-Functional Proteomics for the Analysis of Immune Regulation. In: Lazar, I., Kontoyianni, M., Lazar, A. (eds) Proteomics for Drug Discovery. Methods in Molecular Biology, vol 1647. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7201-2_9

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  • DOI: https://doi.org/10.1007/978-1-4939-7201-2_9

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7200-5

  • Online ISBN: 978-1-4939-7201-2

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