Analysis of Histone Deacetylases Sumoylation by Immunoprecipitation Techniques

Methods Mol Biol. 2017:1510:339-351. doi: 10.1007/978-1-4939-6527-4_25.

Abstract

Histone deacetylases (HDACs) are controlling dynamic protein acetylation by removing acetyl moieties from lysine. Histone deacetylases themselves are regulated on the posttranslational level, including modifications with small ubiquitin-like modifier (SUMO) proteins. Detecting SUMO modifications of deacetylases by immunoblotting is technically challenging due to the typically low ratio of the modified compared to the unmodified species. Here, we describe a set of methods for the detection of endogenous sumoylated HDACs by immunoprecipitation and immunoblotting techniques.

Keywords: Epigenetics; HDAC; Histone deacetylases; Immunoblotting; Immunoprecipitation; Posttranslational modification; SUMO; Sumoylation.

MeSH terms

  • Blotting, Western / methods
  • Epigenesis, Genetic
  • HEK293 Cells
  • Histone Deacetylase 1 / genetics
  • Histone Deacetylase 1 / metabolism*
  • Humans
  • Immunoprecipitation / methods
  • Lysine / metabolism
  • Protein Processing, Post-Translational*
  • SUMO-1 Protein / genetics
  • SUMO-1 Protein / metabolism*
  • Small Ubiquitin-Related Modifier Proteins / genetics
  • Small Ubiquitin-Related Modifier Proteins / metabolism*
  • Sumoylation
  • Transfection
  • Ubiquitins / genetics
  • Ubiquitins / metabolism*

Substances

  • SUMO-1 Protein
  • SUMO1 protein, human
  • SUMO2 protein, human
  • SUMO3 protein, human
  • Small Ubiquitin-Related Modifier Proteins
  • Ubiquitins
  • HDAC1 protein, human
  • Histone Deacetylase 1
  • Lysine