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Phosphoproteome and Proteome Sample Preparation from Mouse Tissues for Circadian Analysis

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Circadian Clocks

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2130))

Abstract

Recent advances in mass spectrometry (MS)-based quantitative proteomics now allow the identification and quantification of deep proteomes and post-translational modifications (PTMs) in relatively short times. Therefore, in the last few years, this technology has proven successful in the circadian field to characterize temporal oscillations of the proteome and more recently PTMs in cellular systems and in tissues. In this chapter, we describe a robust and simple protocol, based on the EasyPhos workflow, to enable preparation of large number of proteomes and phosphoproteomes from mouse tissues for MS-based quantitative analysis. We additionally discuss computational methods to analyze proteome and phosphoproteome time series to determine circadian oscillations.

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Acknowledgments

We thank Matthias Mann and group members for their support.

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Authors and Affiliations

  1. Department of Proteomics and Signal Transduction, Max-Planck Institute of Biochemistry, Martinsried, Germany

    Franziska Brüning

  2. Institute of Medical Psychology, Faculty of Medicine, LMU, Munich, Germany

    Franziska Brüning & Maria S. Robles

  3. School of Life and Environmental Sciences, University of Sydney, Sydney, NSW, Australia

    Sean J. Humphrey

Authors
  1. Franziska Brüning
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  2. Sean J. Humphrey
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  3. Maria S. Robles
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Corresponding author

Correspondence to Maria S. Robles .

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Editors and Affiliations

  1. Chronobiology and Sleep Research Group, Institute of Pharmacology and Toxicology, University of Zurich, Zürich, Switzerland

    Steven A. Brown

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Brüning, F., Humphrey, S.J., Robles, M.S. (2021). Phosphoproteome and Proteome Sample Preparation from Mouse Tissues for Circadian Analysis. In: Brown, S.A. (eds) Circadian Clocks. Methods in Molecular Biology, vol 2130. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0381-9_14

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  • DOI: https://doi.org/10.1007/978-1-0716-0381-9_14

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0380-2

  • Online ISBN: 978-1-0716-0381-9

  • eBook Packages: Springer Protocols

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