Chemoprobe-based assays of histone lysine demethylase 1A target occupation enable in vivo pharmacokinetics and pharmacodynamics studies of KDM1A inhibitors.

The Journal of Biological Chemistry
Cristina MascaróTamara Maes

Abstract

Screening of cellular activity for inhibitors of histone lysine modifiers is most frequently performed indirectly by analyzing changes in the total levels of histone marks targeted by lysine methylases/demethylases. However, inhibition of histone lysine modifiers often leads to local rather than total changes in histone marks. Also, because histone modifications can be modulated by more than one cellular enzyme, it is not always clear whether changes in histone marks are a direct or indirect consequence of the inhibitor treatment applied. Direct assessment of target occupation can provide a useful tool to quantify the fraction of an epigenetic modifier that is bound to a pharmacological inhibitor (target engagement). Here, we developed and used a novel chemoprobe-based immunoassay to quantify target engagement in cells. Quantification of the fraction of free KDM1A was made possible, in an immune-based assay, by coupling a biotinylated chemoprobe to a warhead capable of selectively and irreversibly binding to the free active form of KDM1A. The results obtained confirmed that this approach is able to determine the degree of target engagement in a dose-dependent manner. Furthermore, the assay can be also used on tissue extracts to...Continue Reading

References

Nov 30, 2002·The Journal of Biological Chemistry·Manuela CervelliPaolo Mariottini
Oct 15, 2005·The Journal of Biological Chemistry·Federico FornerisAndrea Mattevi
Jun 24, 2006·Chemistry & Biology·Min Gyu LeeRamin Shiekhattar
Jan 18, 2007·Proceedings of the National Academy of Sciences of the United States of America·Cleo M Salisbury, Benjamin F Cravatt
Jan 25, 2011·Nature Biotechnology·Marcus BantscheffGerard Drewes
Aug 12, 2014·Epigenomics·Peter Birk Rasmussen, Peter Staller
Dec 19, 2016·Bioorganic & Medicinal Chemistry·Maria E OurailidouFrank J Dekker

❮ Previous
Next ❯

Citations

Jul 23, 2019·Chemical Communications : Chem Comm·James E Kath, Aleksandra Baranczak
Aug 14, 2020·Chembiochem : a European Journal of Chemical Biology·Jenna N BeyerGeorge M Burslem
May 19, 2019·The Journal of Biological Chemistry·Aseem Z Ansari
Sep 6, 2020·Bioorganic Chemistry·Gang JiaZhiyu Song
Dec 21, 2021·ACS Pharmacology & Translational Science·Natalia SacilottoRobert Soliva

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.