Detection and quantification of histone methyltransferase activity in vitro

Nwamaka J Idigo, Philipp Voigt

Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)peer-review

Abstract

Histone methyltransferases (HMTs) catalyze the methylation of lysine and arginine residues in histone as well as nonhistone substrates. In vitro histone methyltransferase assays have been instrumental in identifying HMTs, and they continue to be invaluable tools for the study of these important enzymes, revealing novel substrates and modes of regulation.Here we describe a universal protocol to examine HMT activity in vitro that can be adapted to a range of HMTs, substrates, and experimental objectives. We provide protocols for the detection of activity based on incorporation of 3H-labeled methyl groups from S-adenosylmethionine (SAM), methylation-specific antibodies, and quantification of the reaction product S-adenosylhomocysteine (SAH).

Original languageEnglish
Title of host publicationHistone methyltransferases
Subtitle of host publicationMethods and protocols
Pages43-61
Number of pages19
Volume2529
ISBN (Electronic)978-1-0716-2481-4
DOIs
Publication statusE-pub ahead of print - 23 Jun 2022

Publication series

NameMethods in molecular biology (Clifton, N.J.)
PublisherHumana Press
ISSN (Print)1064-3745

Keywords

  • histone methyltransferases
  • histone-lysine N-methyltransferase
  • histones
  • methylation
  • protein processing
  • S-Adenosylmethionine
  • post-translational

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