Cancer-associated 2-oxoglutarate analogues modify histone methylation by inhibiting histone lysine demethylases

J Mol Biol. 2018 Sep 14;430(18 Pt B):3081-3092. doi: 10.1016/j.jmb.2018.06.048. Epub 2018 Jul 5.


Histone lysine demethylases (KDMs) are 2-oxoglutarate-dependent dioxygenases (2-OGDDs) that regulate gene expression by altering chromatin structure. Their dysregulation has been associated with many cancers. We set out to study the catalytic and inhibitory properties of human KDM4A, KDM4B, KDM5B, KDM6A and KDM6B, aiming in particular to reveal which of these enzymes are targeted by cancer-associated 2-oxoglutarate (2-OG) analogues. We used affinity-purified insect cell-produced enzymes and synthetic peptides with trimethylated lysines as substrates for the in vitro enzyme activity assays. In addition, we treated breast cancer cell lines with cell-permeable forms of 2-OG analogues and studied their effects on the global histone methylation state. Our data show that KDMs have substrate specificity. Among the enzymes studied, KDM5B had the highest affinity for the peptide substrate but the lowest affinity for the 2-OG and the Fe2+ cosubstrate/cofactors. R-2-hydroxyglutarate (R-2HG) was the most efficient inhibitor of KDM6A, KDM4A and KDM4B, followed by S-2HG. This finding was supported by accumulations of the histone H3K9me3 and H3K27me3 marks in cells treated with the cell-permeable forms of these compounds. KDM5B was especially resistant to inhibition by R-2HG, while citrate was the most efficient inhibitor of KDM6B. We conclude that KDM catalytic activity is susceptible to inhibition by tumorigenic 2-OG analogues and suggest that the inhibition of KDMs is involved in the disease mechanism of cancers in which these compounds accumulate, such as the isocitrate dehydrogenase mutations.

Keywords: KDM; R-2HG; S-2HG; fumarate; succinate.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Enzyme Activation
  • Histone Demethylases / antagonists & inhibitors*
  • Histone Demethylases / metabolism*
  • Histones / metabolism*
  • Humans
  • Ketoglutaric Acids / chemistry
  • Ketoglutaric Acids / pharmacology*
  • Kinetics
  • Methylation / drug effects
  • Substrate Specificity


  • Histones
  • Ketoglutaric Acids
  • Histone Demethylases