A protocol for high-throughput screening of histone lysine demethylase 4 inhibitors using TR-FRET assay

STAR Protoc. 2021 Aug 26;2(3):100702. doi: 10.1016/j.xpro.2021.100702. eCollection 2021 Sep 17.

Abstract

Identification of diverse chemotypes of selective KDM4 inhibitors is important for exploring and validating the roles of KDM4s in the pathogenesis of human disease and for developing therapies. Here, we report a protocol for high-throughput screening of KDM4 inhibitors using TR-FRET demethylation functional assay. We describe this protocol for screen of KDM4B inhibitors, which can be modified to screen inhibitors of other JmjC-domain-containing KDMs. For complete details on the use and execution of this protocol, please refer to Singh et al. (2021).

Keywords: Cancer; Chemistry; High Throughput Screening; Molecular/Chemical Probes; Protein Biochemistry; Protein expression and purification; Structural Biology.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Drug Evaluation, Preclinical / methods*
  • Fluorescence Resonance Energy Transfer / methods*
  • High-Throughput Screening Assays / methods*
  • Histone Demethylases / antagonists & inhibitors
  • Histones / metabolism
  • Humans
  • Jumonji Domain-Containing Histone Demethylases / antagonists & inhibitors
  • Jumonji Domain-Containing Histone Demethylases / metabolism
  • Lysine
  • Substrate Specificity

Substances

  • Histones
  • Histone Demethylases
  • Jumonji Domain-Containing Histone Demethylases
  • KDM4B protein, human
  • Lysine