Purification of Histone Variant-Interacting Chaperone Complexes.

Abstract

Identification of the interaction partners of a protein is one of useful straightforward methods to gain insight into the molecular functions of the protein in cells. The pre-deposited forms of histones are associated with the specific histone chaperones to assemble into chromatin. Here, I describe an affinity purification method using the FLAG/HA double epitope-tagging technique and its application to purify particular histone variant-interacting chaperone complexes from soluble fraction to study dynamic chromatin functions. The purification is performed under low salt condition to obtain native histone variant complexes, and it would be useful to identify the specific chaperone proteins involved in the specific chromatin functions via histone variants.