5V7T

crystal structure of PARP14 bound to N-{4-[4-(diphenylmethoxy)piperidin-1-yl]butyl}[1,2,4]triazolo[4,3-b]pyridazin-6-amine inhibitor

Summary for 5V7T

• Entry DOI: 10.2210/pdb5v7t/pdb
• Related: 5V7W
• Descriptor: Poly [ADP-ribose] polymerase 14, N-{4-[4-(diphenylmethoxy)piperidin-1-yl]butyl}[1,2,4]triazolo[4,3-b]pyridazin-6-amine (3 entities in total)
• Functional Keywords: human parp14, transferase-transferase inhibitor complex, transferase/transferase inhibitor
• Biological source: Homo sapiens (Human)
• Cellular location: Nucleus: Q460N5
• Total number of polymer chains: 1
• Total formula weight: 22558.08

Authors

saikatendu, k.s.,Hirozane, M. (deposition date: 2017-03-20, release date: 2017-05-03, Last modification date: 2024-11-06)

Primary citation

Yoneyama-Hirozane, M.,Matsumoto, S.I.,Toyoda, Y.,Saikatendu, K.S.,Zama, Y.,Yonemori, K.,Oonishi, M.,Ishii, T.,Kawamoto, T.
Identification of PARP14 inhibitors using novel methods for detecting auto-ribosylation.
Biochem. Biophys. Res. Commun., 486:626-631, 2017

PubMed Abstract: Poly(ADP-ribose) polymerases (PARPs) use nicotinamide adenine dinucleotide (NAD) as a co-substrate to transfer ADP-ribose when it releases nicotinamide as the metabolized product. Enzymes of the PARP family play key roles in detecting and repairing DNA, modifying chromatin, regulating transcription, controlling energy metabolism, and inducing cell death. PARP14, the original member of the PARP family, has been reported to be associated with the development of inflammatory diseases and various cancer types, making it a potential therapeutic target. In this study, we purified the macrodomain-containing PARP14 enzyme and established an assay for detecting the auto-ribosylation activity of PARP14 using RapidFire high-throughput mass spectrometry and immunoradiometric assay using [H]NAD. Subsequently, we performed high-throughput screening using the assays and identified small-molecule hit compounds, which showed NAD-competitive and PARP14-selective inhibitory activities. Co-crystal structures of PARP14 with certain hit compounds revealed that the inhibitors bind to the NAD-binding site. Finally, we confirmed that the hit compounds interacted with intracellular PARP14 by a cell-based protein stabilization assay. Thus, we successfully identified primary candidate compounds for further investigation.

PubMed: 28315326
DOI: 10.1016/j.bbrc.2017.03.052

Experimental method

X-RAY DIFFRACTION (2.3 Å)