A liquid chromatography/mass spectrometry-based generic detection method for biochemical assay and hit discovery of histone methyltransferases

Anal Biochem. 2013 Dec 15;443(2):214-21. doi: 10.1016/j.ab.2013.08.029. Epub 2013 Sep 7.

Abstract

Epigenetic modifications of the genome, such as DNA methylation and posttranslational modifications of histone proteins, contribute to gene regulation. Growing evidence suggests that histone methyltransferases are associated with the development of various human diseases, including cancer, and are promising drug targets. High-quality generic assays will facilitate drug discovery efforts in this area. In this article, we present a liquid chromatography/mass spectrometry (LC/MS)-based S-adenosyl homocysteine (SAH) detection assay for histone methyltransferases (HMTs) and its applications in HMT drug discovery, including analyzing the activity of newly produced enzymes, developing and optimizing assays, performing focused compound library screens and orthogonal assays for hit confirmations, selectivity profiling against a panel of HMTs, and studying mode of action of select hits. This LC/MS-based generic assay has become a critical platform for our methyltransferase drug discovery efforts.

Keywords: Assay; Histone; LC/MS; Methyltransferase; S-Adenosyl homocysteine.

MeSH terms

  • Chromatography, Liquid / methods*
  • Drug Discovery / methods*
  • Enzyme Assays / methods
  • Histone Methyltransferases
  • Histone-Lysine N-Methyltransferase / analysis*
  • Histone-Lysine N-Methyltransferase / metabolism
  • Humans
  • S-Adenosylhomocysteine / analysis
  • S-Adenosylhomocysteine / metabolism
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*

Substances

  • S-Adenosylhomocysteine
  • Histone Methyltransferases
  • Histone-Lysine N-Methyltransferase