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SILAC-Based Quantification of Sirt1-Responsive Lysine Acetylome

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Book cover Sirtuins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1077))

Abstract

Stable Isotope Labeling by Amino acids in Cell culture (SILAC) is one of the in vivo metabolic labeling methods widely used for dynamic analysis of protein modifications. Here, we describe a general approach to applying SILAC, in combination with affinity enrichment of acetyllysine peptides and mass spectrometry, to study the dynamic changes of the Lysine acetylome in response to Sirt1. The method should be applicable to quantify changes to other post translational modifications in diverse cellular systems.

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Acknowledgements

Research reported in this publication was supported by the National Cancer Institute of the National Institutes of Health under Award Number CA126832 and RR020839 to Y. Zhao. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Y. Zhao is also supported by the Nancy and Leonard Florsheim Family Fund. Y. Chen is supported by NIH Fellowship (T32 HL094282).

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Authors and Affiliations

  1. Ben May Department for Cancer Research, University of Chicago, Chicago, IL, USA

    Yue Chen, Gozde Colak & Yingming Zhao

Authors
  1. Yue Chen
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  2. Gozde Colak
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  3. Yingming Zhao
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Editor information

Editors and Affiliations

  1. Duke University Medical Center Institute of Molecular Physiology, Durham, North Carolina, USA

    Matthew D. Hirschey

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Chen, Y., Colak, G., Zhao, Y. (2013). SILAC-Based Quantification of Sirt1-Responsive Lysine Acetylome. In: Hirschey, M. (eds) Sirtuins. Methods in Molecular Biology, vol 1077. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-637-5_7

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  • DOI: https://doi.org/10.1007/978-1-62703-637-5_7

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-636-8

  • Online ISBN: 978-1-62703-637-5

  • eBook Packages: Springer Protocols

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