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Global Profiling of Sirtuin Deacylase Substrates Using a Chemical Proteomic Strategy and Validation by Fluorescent Labeling

  • Shuai Zhang
  • Nicole A. Spiegelman
  • Hening LinEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2009)

Abstract

Protein fatty-acylation is an important posttranslational modification (PTM) and has been associated with many fundamental biological processes. Sirtuins, the nicotinamide adenine dinucleotide (NAD)-dependent class of histone deacetylases have been reported to possess lysine defatty-acylase activity. Comprehensive substrate profiling of sirtuins will help to establish the function of both protein lysine fatty acylation and its regulation by sirtuins. Here, we describe a chemical proteomic strategy to globally profile sirtuin defatty-acylation substrates and a fluorescent labeling method to validate sirtuin substrates.

Key words

Protein fatty acylation Sirtuin substrates Proteomic profiling SILAC Fluorescent labeling 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Shuai Zhang
    • 1
  • Nicole A. Spiegelman
    • 1
  • Hening Lin
    • 2
    Email author
  1. 1.Department of Chemistry and Chemical BiologyCornell UniversityIthacaUSA
  2. 2.Department of Chemistry and Chemical Biology, Howard Hughes Medical InstituteCornell UniversityIthacaUSA

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