A FRET-based assay for screening SIRT6 modulators

Eur J Med Chem. 2015:96:245-9. doi: 10.1016/j.ejmech.2015.04.008. Epub 2015 Apr 9.

Abstract

SIRT6, as one of these seven sirtuins, has been shown to have the therapeutic potentials for treating several human diseases. A fluorogenic assay for SIRT6 has been developed to screen their small molecule modulators based on the recent discovery that SIRT6 is a defatty-acylase (removing long chain fatty acyl groups). However, this assay uses a fluorogenic peptide containing 7-amino-4-methylcoumarin (AMC), which becomes the cause of false positive hits from screenings. To overcome this, we have developed an alternative method called a FRET-based assay suitable for screening SIRT6 modulators, which will be reliable and useful in a high-throughput format since no AMC group present in this assay.

Keywords: Deacetylation; Defatty-acylation; FRET assay; Fluorogenic assay; Sirtuin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Coumarins / chemical synthesis
  • Coumarins / chemistry
  • Coumarins / pharmacology*
  • Dose-Response Relationship, Drug
  • Drug Evaluation, Preclinical*
  • Enzyme Inhibitors / chemical synthesis
  • Enzyme Inhibitors / chemistry
  • Enzyme Inhibitors / pharmacology*
  • Fluorescence Resonance Energy Transfer*
  • Fluorescent Dyes / analysis
  • Fluorescent Dyes / chemical synthesis
  • Fluorescent Dyes / chemistry
  • Humans
  • Molecular Structure
  • Sirtuins / antagonists & inhibitors*
  • Sirtuins / metabolism
  • Structure-Activity Relationship

Substances

  • Coumarins
  • Enzyme Inhibitors
  • Fluorescent Dyes
  • SIRT6 protein, human
  • Sirtuins
  • 7-amino-4-methylcoumarin