Chemical tools for profiling the intracellular ADP-ribosylated proteome.

Autor:	Draganov SD; Department of Chemistry, Molecular Sciences Research Hub, Imperial College London London W12 0BZ UK e.tate@imperial.ac.uk., Gruet MJ; Department of Chemistry, Molecular Sciences Research Hub, Imperial College London London W12 0BZ UK e.tate@imperial.ac.uk.; Department of Surgery and Cancer, Institute of Reproductive and Developmental Biology, Imperial College London London W12 0HS UK., Conole D; Department of Chemistry, Molecular Sciences Research Hub, Imperial College London London W12 0BZ UK e.tate@imperial.ac.uk., Balcells C; Department of Surgery and Cancer, Institute of Reproductive and Developmental Biology, Imperial College London London W12 0HS UK., Siskos AP; Department of Surgery and Cancer, Institute of Reproductive and Developmental Biology, Imperial College London London W12 0HS UK., Keun HC; Department of Surgery and Cancer, Institute of Reproductive and Developmental Biology, Imperial College London London W12 0HS UK., Haskard DO; Faculty of Medicine, National Heart and Lung Institute, Imperial Centre for Translational and Experimental Medicine, Imperial College London London W12 0HS UK., Tate EW; Department of Chemistry, Molecular Sciences Research Hub, Imperial College London London W12 0BZ UK e.tate@imperial.ac.uk.
Jazyk:	angličtina
Zdroj:	RSC chemical biology [RSC Chem Biol] 2024 May 22; Vol. 5 (7), pp. 640-651. Date of Electronic Publication: 2024 May 22 (Print Publication: 2024).
DOI:	10.1039/d4cb00043a
Abstrakt:	The post-translational modification (PTM) ADP-ribosylation plays an important role in cell signalling and regulating protein function and has been implicated in the development of multiple diseases, including breast and ovarian cancers. Studying the underlying mechanisms through which this PTM contributes towards disease development, however, has been hampered by the lack of appropriate tools for reliable identification of physiologically relevant ADP-ribosylated proteins in a live-cell environment. Herein, we explore the application of an alkyne-tagged proprobe, 6Yn-ProTide-Ad (6Yn-Pro) as a chemical tool for the identification of intracellular ADP-ribosylated proteins through metabolic labelling. We applied targeted metabolomics and chemical proteomics in HEK293T cells treated with 6Yn-Pro to demonstrate intracellular metabolic conversion of the probe into ADP-ribosylation cofactor 6Yn-NAD + , and subsequent labelling and enrichment of PARP1 and multiple known ADP-ribosylated proteins in cells under hydrogen peroxide-induced stress. We anticipate that the approach and methodology described here will be useful for future identification of novel intracellular ADP-ribosylated proteins. Competing Interests: The authors declare no conflict of interest. (This journal is © The Royal Society of Chemistry.)
Databáze:	MEDLINE
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