Methods and tips for the purification of human histone methyltransferases

Recently developed biochemical techniques have enabled researchers to study historic modifications more easily and accurately. One of these triodifications, historic lysine methylation, has been shown to be highly stable and to represent an epigenetic alteration. Extensive biochemical analyses have led to discoveries about the nature and functions of this modification, thus accelerating our understanding of this crucial epigenctic event. Here we describe basic methods for purification and biochemical analysis of lysine-directed, historic methyltransferases from HeLa cell-derived extracts. In the section on substrate preparation, we describe a simple method for the preparation of recombinant substrates, although we recommend using native substrates for initial detection of the activities. The purification protocols for several histone methyltransferases have been streamlined so that those researchers with a basic understanding of biochemistry can perform them. We also describe many tips and provide suggestions to avoid common pitfalls in the biochemical analysis of histone methyltransferases. (C) 2003 Elsevier Science (USA). All rights reserved.