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Related Experiment Video

Updated: Aug 25, 2025

Single-Step Enrichment of a TAP-Tagged Histone Deacetylase of the Filamentous Fungus Aspergillus nidulans for Enzymatic Activity Assay
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Continuous Histone Deacylase Activity Assays.

Matthes Zessin1, Marat Meleshin2, Wolfgang Sippl1

  • 1Department of Medicinal Chemistry, Institute of Pharmacy, Martin-Luther-University Halle-Wittenberg, Halle, Saale, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|October 18, 2022
PubMed
Summary

New assays monitor histone deacetylase (HDAC) activity for drug discovery. These methods track Zn2+-dependent HDACs and NAD+-dependent sirtuins, aiding the development of novel therapeutics.

Keywords:
Activity assayHistone deacetylaseInhibitor measurementSirtuin

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Drug Discovery

Background:

  • Protein lysine acylation is a common post-translational modification.
  • Histone deacetylases (HDACs) regulate critical cellular processes including metabolism, aging, and immune response.
  • HDAC modulators are valuable in drug discovery, necessitating robust assay development.

Purpose of the Study:

  • To develop direct and continuous assay formats for monitoring HDAC enzymatic activity.
  • To enable the study of both Zn2+-dependent HDACs and NAD+-dependent sirtuins.
  • To facilitate efficient screening of HDAC inhibitors.

Main Methods:

  • UV-Vis-spectroscopy for monitoring Zn2+-dependent HDACs.
  • Fluorescence-based assay formats for NAD+-dependent HDACs (sirtuins).
  • Fluorescence displacement assays for screening HDAC inhibitors.

Main Results:

  • Demonstration of UV-Vis-spectroscopy for tracking Zn2+-dependent HDAC activity.
  • Establishment of fluorescence-based assays for sirtuin activity monitoring.
  • Development of efficient fluorescence displacement assays for HDAC inhibitor screening.

Conclusions:

  • The developed assay formats provide essential tools for studying HDACs and sirtuins.
  • These assays support drug discovery efforts targeting enzymes involved in various diseases.
  • Continuous monitoring and inhibitor screening capabilities are crucial for advancing therapeutic development.