Highly Sensitive Lysine Deacetylase Assay Based on Acetylated Firefly Luciferase

Biochemistry
Martin SpinckHeinz Neumann

Abstract

Lysine deacetylases (KDACs) play important roles in many physiological processes and are implicated in many human diseases. Hence, the search for modulators of KDACs is very active, and reliable assays for monitoring their activity are key to success. Here, we describe a new KDAC assay based on Firefly luciferase harboring an acetylation on an essential active site lysine. We show that several KDACs can reverse this modification and hence activate luciferase. This new assay is extremely sensitive, reliable, and fast and can be performed in a continuous format. We used this assay to screen a small library of compounds and identified several novel effectors of SirT2 with low micromolar activity.

References

Nov 30, 2000·Biochemical and Biophysical Research Communications·J LandryR Sternglanz
Sep 26, 2002·The Journal of Biological Chemistry·Kevin J BittermanDavid A Sinclair
Feb 8, 2003·Chemistry & Biology·Dennis WegenerAndreas Schwienhorst
Aug 27, 2003·Nature·Konrad T HowitzDavid A Sinclair
Aug 1, 1953·The Biochemical Journal·M DIXON
Oct 3, 2003·The Journal of Biological Chemistry·Michael D JacksonJohn M Denu
Feb 3, 2005·The Journal of Biological Chemistry·Matt KaeberleinBrian K Kennedy
Mar 8, 2005·The Journal of Biological Chemistry·Margie T BorraJohn M Denu
Dec 8, 2006·Journal of Medicinal Chemistry·Johannes TrappManfred Jung
Jun 26, 2007·Science·Tiago Fleming OuteiroAleksey G Kazantsev
Jun 27, 2007·Nature Biotechnology·Kaihang WangJason W Chin
Jul 21, 2009·Analytical Biochemistry·Brian C SmithJohn M Denu
Jan 12, 2010·The Journal of Biological Chemistry·Michelle PacholecKay Ahn
Sep 14, 2011·Journal of Biomolecular Screening·Peter T RyeWilliam A Lamarr
Jun 13, 2012·Angewandte Chemie·Chan Hyuk KimPeter G Schultz
Oct 23, 2012·The Journal of Biological Chemistry·Carlos SebastiánRaul Mostoslavsky
Nov 28, 2012·PloS One·Melanie GertzClemens Steegborn
Nov 30, 2012·Chemical Communications : Chem Comm·Michael J GattnerThomas Carell
Jan 3, 2014·Expert Opinion on Drug Discovery·Mike SchutkowskiClemens Steegborn
Jul 24, 2014·Nature Reviews. Molecular Cell Biology·Chunaram ChoudharyMatthias Mann
Jan 16, 2015·ACS Chemical Biology·Bryan J WilkinsHeinz Neumann
Apr 25, 2015·ACS Chemical Biology·Han XiaoPeter G Schultz
May 23, 2015·Organic & Biomolecular Chemistry·William A Knight, T Ashton Cropp
Oct 13, 2015·Biochemical and Biophysical Research Communications·Yan LiBin He
Mar 5, 2016·Scientific Reports·Sabine SchusterMike Schutkowski
May 13, 2017·Proceedings of the Japan Academy. Series B, Physical and Biological Sciences·Minoru YoshidaAkihiro Ito

❮ Previous
Next ❯

Citations

Mar 19, 2020·Angewandte Chemie·Martin SpinckHeinz Neumann
Apr 21, 2021·Journal of Medicinal Chemistry·Martin SpinckHeinz Neumann
Oct 30, 2021·Advanced Biology·Petra Neumann-StaubitzHeinz Neumann
Nov 9, 2021·Current Protocols·Leonie G GrafMichael Lammers
Dec 15, 2021·Chembiochem : a European Journal of Chemical Biology·Ruotong XiaoWeimin Xuan

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.