University of Oulu

Haikarainen, T., Maksimainen, M. M., Obaji, E., & Lehtiö, L. (2018). Development of an Inhibitor Screening Assay for Mono-ADP-Ribosyl Hydrolyzing Macrodomains Using AlphaScreen Technology. SLAS DISCOVERY: Advancing Life Sciences R&D, 23(3), 255–263.

Development of an inhibitor screening assay for mono-ADP-ribosyl hydrolyzing macrodomains using AlphaScreen technology

Saved in:
Author: Haikarainen, Teemu1; Maksimainen, Mirko M.1; Obaji, Ezeogo1;
Organizations: 1Faculty of Biochemistry and Molecular Medicine and Biocenter Oulu, University of Oulu, Oulu, Finland
Format: article
Version: accepted version
Access: open
Online Access: PDF Full Text (PDF, 1 MB)
Persistent link:
Language: English
Published: SAGE Publications, 2018
Publish Date: 2019-02-25


Protein mono-ADP-ribosylation is a posttranslational modification involved in the regulation of several cellular signaling pathways. Cellular ADP-ribosylation is regulated by ADP-ribose hydrolases via a hydrolysis of the protein-linked ADP-ribose. Most of the ADP-ribose hydrolases share a macrodomain fold. Macrodomains have been linked to several diseases, such as cancer, but their cellular roles are mostly unknown. Currently, there are no inhibitors available targeting the mono-ADP-ribose hydrolyzing macrodomains. We have developed a robust AlphaScreen assay for the screening of inhibitors against macrodomains having mono-ADP-ribose hydrolysis activity. We utilized this assay for validatory screening against human MacroD1 and identified five compounds inhibiting the macrodomain. Dose–response measurements and an orthogonal assay further validated four of these compounds as MacroD1 inhibitors. The developed assay is homogenous, easy to execute, and suitable for the screening of large compound libraries. The assay principle can also be adapted for other ADP-ribose hydrolyzing macrodomains, which can utilize a biotin-mono-ADP-ribosylated protein as a substrate.

see all

Series: SLAS discovery
ISSN: 2472-5552
ISSN-E: 2472-5560
ISSN-L: 2472-5552
Volume: 23
Issue: 3
Pages: 255 - 263
DOI: 10.1177/2472555217737006
Type of Publication: A1 Journal article – refereed
Field of Science: 1182 Biochemistry, cell and molecular biology
Funding: This work was funded by Biocenter Oulu, Orion research foundation, and Academy of Finland (grant No. 287063 and 294085 for LL and grant No. 266922 for TH).
Academy of Finland Grant Number: 287063
Detailed Information: 287063 (Academy of Finland Funding decision)
294085 (Academy of Finland Funding decision)
Copyright information: © 2017 SAGE Publications. Reprinted by permission of SAGE Publications.