title: Detection of Sumo Modification of Endogenous Histone Deacetylase 2 (HDAC2) in Mammalian Cells.
abstract: Small ubiquitin-related modifier (SUMO) is an ubiquitin-like protein that is covalently attached to a variety of target proteins and has a significant role in their regulation. HDAC2 is an important epigenetic regulator, promoting the deacetylation of histones and non-histone proteins. HDAC2 has been shown to be modified by SUMO1 at lysine 462. Here we describe how to detect SUMO modification of endogenous HDAC2 in mammalian cells by immunoblotting. Although in this chapter we use this method to detect HDAC2 modification in mammalian cells, this protocol can be used for any cell type or for any protein of interest.
authors.affiliation: Department of Experimental Oncology, European Institute of Oncology, IFOM-IEO Campus, 20139, Milan, Italy.
authors.forename: Simona
authors.initials: S
authors.lastname: Citro
authors.affiliation: Department of Experimental Oncology, European Institute of Oncology, IFOM-IEO Campus, 20139, Milan, Italy. susanna.chiocca@ieo.eu.
authors.forename: Susanna
authors.initials: S
authors.lastname: Chiocca
mesh_terms: D045744:Cell Line, Tumor; D044127:Epigenesis, Genetic; D006367:HeLa Cells; D056464:Histone Deacetylase 2 / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D008239:Lysine / Q000378:metabolism*; D061986:MCF-7 Cells; D011499:Protein Processing, Post-Translational; D025841:Small Ubiquitin-Related Modifier Proteins / Q000378:metabolism*; D058207:Sumoylation
chemical_list: D025841:Small Ubiquitin-Related Modifier Proteins; C533423:HDAC2 protein, human; D056464:Histone Deacetylase 2; D008239:Lysine
keywords: HDAC2; Immunoblotting; PTM; Posttranslational modification; SUMO; UBC9