Isolation and characterization of histone H3 lysine 4 demethylase-containing complexes.
Histone methylation is involved in the regulation of many cellular processes. In the past 2 years, several histone demethylases including BHC110/LSD1 have been characterized. BHC110, the first known histone lysine demethylase, removes methyl groups from methylated histone H3 lysine 4 and has been found in many multi-protein complexes. Using one-step affinity purification, we have isolated enzymatically active BHC110-containing complexes. Here, we detail the methods used for the isolation and characterization of these histone demethylase complexes from a human stable cell line.
Lee
Min Gyu
MG
The Wistar Institute, 3601 Spruce Street, Philadelphia, PA 19104, USA.
Wynder
Christopher
C
Norman
Jessica
J
Shiekhattar
Ramin
R
D002460:Cell Line; D002846:Chromatography, Affinity / Q000379:methods*; D006657:Histones / Q000378:metabolism*; D006801:Humans; D008239:Lysine / Q000378:metabolism; D008745:Methylation; D008780:Methyltransferases / Q000302:isolation & purification* / Q000378:metabolism; D046912:Multiprotein Complexes / Q000302:isolation & purification* / Q000378:metabolism; D010089:Oxidoreductases, N-Demethylating / Q000302:isolation & purification* / Q000378:metabolism; D014162:Transfection / Q000379:methods
D006657:Histones; D046912:Multiprotein Complexes; D010089:Oxidoreductases, N-Demethylating; D008780:Methyltransferases; D008239:Lysine