A new split-luciferase complementation assay identifies pentachlorophenol as an inhibitor of apoptosome formation.

Autor: Tashakor A; Pharmacology and Therapeutics, School of Medicine, NUI Galway, Ireland., H-Dehkordi M; Pharmacology and Therapeutics, School of Medicine, NUI Galway, Ireland., O'Connell E; Genomics and Screening Core, National Centre for Biomedical Engineering Science, NUI Galway, Ireland., Gomez Ganau S; PROTOQSAR, Valencia, Spain., Gozalbes R; PROTOQSAR, Valencia, Spain., Eriksson LA; University of Gothenburg, Sweden., Hosseinkhani S; Tarbiat Modares University, Tehran, Iran., Fearnhead HO; Pharmacology and Therapeutics, School of Medicine, NUI Galway, Ireland.
Jazyk: angličtina
Zdroj: FEBS open bio [FEBS Open Bio] 2019 Jul; Vol. 9 (7), pp. 1194-1203. Date of Electronic Publication: 2019 May 29.
DOI: 10.1002/2211-5463.12646
Abstrakt: The expense and time required for in vivo reproductive and developmental toxicity studies have driven the development of in vitro alternatives. Here, we used a new in vitro split luciferase-based assay to screen a library of 177 toxicants for inhibitors of apoptosome formation. The apoptosome contains seven Apoptotic Protease-Activating Factor-1 (Apaf-1) molecules and induces cell death by activating caspase-9. Apaf-1-dependent caspase activation also plays an important role in CNS development and spermatogenesis. In the in vitro assay, Apaf-1 fused to an N-terminal fragment of luciferase binds to Apaf-1 fused to a C-terminal fragment of luciferase and reconstitutes luciferase activity. Our assay indicated that pentachlorophenol (PCP) inhibits apoptosome formation, and further investigation revealed that PCP binds to cytochrome c. PCP is a wood preservative that reduces male fertility by ill-defined mechanisms. Although the data show that PCP inhibited apoptosome formation, the concentration required suggests that other mechanisms may be more important for PCP's effects on spermatogenesis. Nonetheless, the data demonstrate the utility of the new assay in identifying apoptosome inhibitors, and we suggest that the assay may be useful in screening for reproductive and developmental toxicants.
(© 2019 The Authors. Published by FEBS Press and John Wiley & Sons Ltd.)
Databáze: MEDLINE
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