Identification of Poly(ADP-Ribose) Polymerase Macrodomain Inhibitors Using an AlphaScreen Protocol
Ekblad, Torun ; Verheugd, Patricia ; Lindgren, Anders E ; Nyman, Tomas ; Elofsson, Mikael and Schüler, Herwig LU
(2018)
In SLAS Discovery
23(4).
p.353-362
- Abstract
Macrodomains recognize intracellular adenosine diphosphate (ADP)-ribosylation resulting in either removal of the modification or a protein interaction event. Research into compounds that modulate macrodomain functions could make important contributions. We investigated the interactions of all seven individual macrodomains of the human poly(ADP-ribose) polymerase (PARP) family members PARP9, PARP14, and PARP15 with five mono-ADP-ribosylated (automodified) ADP-ribosyltransferase domains using an AlphaScreen assay. Several mono-ADP-ribosylation-dependent interactions were identified, and they were found to be in the micromolar affinity range using surface plasmon resonance (SPR). We then focused on the interaction between PARP14... (More)
Macrodomains recognize intracellular adenosine diphosphate (ADP)-ribosylation resulting in either removal of the modification or a protein interaction event. Research into compounds that modulate macrodomain functions could make important contributions. We investigated the interactions of all seven individual macrodomains of the human poly(ADP-ribose) polymerase (PARP) family members PARP9, PARP14, and PARP15 with five mono-ADP-ribosylated (automodified) ADP-ribosyltransferase domains using an AlphaScreen assay. Several mono-ADP-ribosylation-dependent interactions were identified, and they were found to be in the micromolar affinity range using surface plasmon resonance (SPR). We then focused on the interaction between PARP14 macrodomain-2 and the mono-ADP-ribosylated PARP10 catalytic domain, and probed a ~1500-compound diverse library for inhibitors of this interaction using AlphaScreen. Initial hit compounds were verified by concentration-response experiments using AlphaScreen and SPR, and they were tested against PARP14 macrodomain-2 and -3. Two initial hit compounds and one chemical analog each were further characterized using SPR and microscale thermophoresis. In conclusion, our results reveal novel macrodomain interactions and establish protocols for identification of inhibitors of such interactions.
(Less)
- author
- Ekblad, Torun
; Verheugd, Patricia
; Lindgren, Anders E
; Nyman, Tomas
; Elofsson, Mikael
and Schüler, Herwig
LU
- publishing date
- 2018-04
- type
- Contribution to journal
- publication status
- published
- keywords
- ADP Ribose Transferases/metabolism, ADP-Ribosylation/drug effects, Adenosine Diphosphate Ribose/metabolism, Biological Assay/methods, Humans, Pentosyltransferases, Poly(ADP-ribose) Polymerase Inhibitors/pharmacology, Poly(ADP-ribose) Polymerases/metabolism
- in
- SLAS Discovery
- volume
- 23
- issue
- 4
- pages
- 10 pages
- publisher
- Society for Laboratory Automation and Screening (SLAS)
- external identifiers
-
- scopus:85044350008
- pmid:29316839
- ISSN
- 2472-5552
- DOI
- 10.1177/2472555217750870
- language
- English
- LU publication?
- no
- id
- 77e3953c-21b0-4f5f-9a56-69eabb506a31
- date added to LUP
- 2024-11-21 17:50:27
- date last changed
- 2025-04-11 21:49:20
@article{77e3953c-21b0-4f5f-9a56-69eabb506a31,
abstract = {{<p>Macrodomains recognize intracellular adenosine diphosphate (ADP)-ribosylation resulting in either removal of the modification or a protein interaction event. Research into compounds that modulate macrodomain functions could make important contributions. We investigated the interactions of all seven individual macrodomains of the human poly(ADP-ribose) polymerase (PARP) family members PARP9, PARP14, and PARP15 with five mono-ADP-ribosylated (automodified) ADP-ribosyltransferase domains using an AlphaScreen assay. Several mono-ADP-ribosylation-dependent interactions were identified, and they were found to be in the micromolar affinity range using surface plasmon resonance (SPR). We then focused on the interaction between PARP14 macrodomain-2 and the mono-ADP-ribosylated PARP10 catalytic domain, and probed a ~1500-compound diverse library for inhibitors of this interaction using AlphaScreen. Initial hit compounds were verified by concentration-response experiments using AlphaScreen and SPR, and they were tested against PARP14 macrodomain-2 and -3. Two initial hit compounds and one chemical analog each were further characterized using SPR and microscale thermophoresis. In conclusion, our results reveal novel macrodomain interactions and establish protocols for identification of inhibitors of such interactions.</p>}},
author = {{Ekblad, Torun and Verheugd, Patricia and Lindgren, Anders E and Nyman, Tomas and Elofsson, Mikael and Schüler, Herwig}},
issn = {{2472-5552}},
keywords = {{ADP Ribose Transferases/metabolism; ADP-Ribosylation/drug effects; Adenosine Diphosphate Ribose/metabolism; Biological Assay/methods; Humans; Pentosyltransferases; Poly(ADP-ribose) Polymerase Inhibitors/pharmacology; Poly(ADP-ribose) Polymerases/metabolism}},
language = {{eng}},
number = {{4}},
pages = {{353--362}},
publisher = {{Society for Laboratory Automation and Screening (SLAS)}},
series = {{SLAS Discovery}},
title = {{Identification of Poly(ADP-Ribose) Polymerase Macrodomain Inhibitors Using an AlphaScreen Protocol}},
url = {{http://dx.doi.org/10.1177/2472555217750870}},
doi = {{10.1177/2472555217750870}},
volume = {{23}},
year = {{2018}},
}