title: Analysis of Histone Deacetylases Sumoylation by Immunoprecipitation Techniques.
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.
authors.affiliation: Department of Biochemistry, Institute of Biochemistry and Biophysics, CMB - Center for Molecular Biomedicine, Friedrich Schiller University Jena, Hans-Knöll-Str. 2, Jena, 07745, Germany.
authors.forename: Tobias
authors.initials: T
authors.lastname: Wagner
authors.affiliation: Department of Biochemistry, Institute of Biochemistry and Biophysics, CMB - Center for Molecular Biomedicine, Friedrich Schiller University Jena, Hans-Knöll-Str. 2, Jena, 07745, Germany.
authors.forename: Maren
authors.initials: M
authors.lastname: Godmann
authors.affiliation: Department of Biochemistry, Institute of Biochemistry and Biophysics, CMB - Center for Molecular Biomedicine, Friedrich Schiller University Jena, Hans-Knöll-Str. 2, Jena, 07745, Germany. t.heinzel@uni-jena.de.
authors.forename: Thorsten
authors.initials: T
authors.lastname: Heinzel
mesh_terms: D015153:Blotting, Western / Q000379:methods; D044127:Epigenesis, Genetic; D057809:HEK293 Cells; D056284:Histone Deacetylase 1 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods; D008239:Lysine / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D025842:SUMO-1 Protein / Q000235:genetics / Q000378:metabolism*; D025841:Small Ubiquitin-Related Modifier Proteins / Q000235:genetics / Q000378:metabolism*; D058207:Sumoylation; D014162:Transfection; D014452:Ubiquitins / Q000235:genetics / Q000378:metabolism*
chemical_list: D025842:SUMO-1 Protein; C495501:SUMO1 protein, human; C117909:SUMO2 protein, human; C492189:SUMO3 protein, human; D025841:Small Ubiquitin-Related Modifier Proteins; D014452:Ubiquitins; C110550:HDAC1 protein, human; D056284:Histone Deacetylase 1; D008239:Lysine
keywords: Epigenetics; HDAC; Histone deacetylases; Immunoblotting; Immunoprecipitation; Posttranslational modification; SUMO; Sumoylation