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Article
Open AccessCovalent PARylation of DNA base excision repair proteins regulates DNA demethylation
The intracellular ATP-ribosyltransferases PARP1 and PARP2, contribute to DNA base excision repair (BER) and DNA demethylation and have been implicated in epigenetic programming in early mammalian development. ...
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Protocol
Subcellular Quantitation of ADP-Ribosylation by High-Content Microscopy
ADP-ribosylation is a posttranslational modification with many functions ranging from the DNA damage response to transcriptional regulation. While nuclear ADP-ribosylation has been extensively studied in the c...
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Article
Open AccessInterplay between ADP-ribosyltransferases and essential cell signaling pathways controls cellular responses
Signaling cascades provide integrative and interactive frameworks that allow the cell to respond to signals from its environment and/or from within the cell itself. The dynamic regulation of mammalian cell sig...
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Protocol
Investigation of Mitochondrial ADP-Ribosylation Via Immunofluorescence
ADP-ribosylation is a posttranslational protein modification, involved in various cellular processes, ranging from DNA-damage repair to apoptosis. While its function has been studied amply with respect to geno...
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Article
Open AccessEngineering Af1521 improves ADP-ribose binding and identification of ADP-ribosylated proteins
Protein ADP-ribosylation is a reversible post-translational modification that regulates important cellular functions. The identification of modified proteins has proven challenging and has mainly been achieved...
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Article
Open AccessValidation of extracellular ligand–receptor interactions by Flow-TriCEPS
The advent of ligand-based receptor capture methodologies, allows the identification of unknown receptor candidates for orphan extracellular ligands. However, further target validation can be tedious, laboriou...
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Protocol
Mono-ADP-Ribosylhydrolase Assays
Despite substantial progress in ADP-ribosylation research in recent years, the identification of ADP-ribosylated proteins, their ADP-ribose acceptors sites, and the respective writers and erasers remains chall...
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Article
Open AccessProteomic analyses identify ARH3 as a serine mono-ADP-ribosylhydrolase
ADP-ribosylation is a posttranslational modification that exists in monomeric and polymeric forms. Whereas the writers (e.g. ARTD1/PARP1) and erasers (e.g. PARG, ARH3) of poly-ADP-ribosylation (PARylation) are...
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Article
Open AccessEcto-ADP-ribosyltransferase ARTC2.1 functionally modulates FcγR1 and FcγR2B on murine microglia
Mammalian ecto-ADP-ribosyltransferases (ecto-ARTs or also ARTCs) catalyze the ADP-ribosylation of cell surface proteins using extracellular nicotinamide adenine dinucleotide (NAD+) as substrate. By this post-tran...
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Article
ADP-ribose-specific chromatin-affinity purification for investigating genome-wide or locus-specific chromatin ADP-ribosylation
This protocol describes how to investigate genome-wide or locus-specific chromatin ADP-ribosylation using ADPr-ChAP.
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Article
Open AccessEpigenetic regulation of nitric oxide synthase 2, inducible (Nos2) by NLRC4 inflammasomes involves PARP1 cleavage
Nitric oxide synthase 2, inducible (Nos2) expression is necessary for the microbicidal activity of macrophages. However, NOS2 over-activation causes multiple inflammatory disorders, suggesting a tight gene regula...
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Protocol
Identification of ADP-Ribose Acceptor Sites on In Vitro Modified Proteins by Liquid Chromatography–Tandem Mass Spectrometry
Protein ADP-ribosylation is a covalent, reversible posttranslational modification (PTM) catalyzed by ADP-ribosyltransferases (ARTs). Proteins can be either mono- or poly-ADP-ribosylated under a variety of phys...
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Protocol
Proteome-Wide Identification of In Vivo ADP-Ribose Acceptor Sites by Liquid Chromatography–Tandem Mass Spectrometry
ADP-ribosylation is a posttranslational modification (PTM) that affects a variety of cellular processes. In recent years, mass spectrometry (MS)-based proteomics has become a valuable tool for studying ADP-rib...
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Article
Open AccessSeveral posttranslational modifications act in concert to regulate gephyrin scaffolding and GABAergic transmission
GABAA receptors (GABAARs) mediate the majority of fast inhibitory neurotransmission in the brain via synergistic association with the postsynaptic scaffolding protein gephyrin and its interaction partners. Howeve...
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Article
Open AccessProteome-wide identification of the endogenous ADP-ribosylome of mammalian cells and tissue
Although protein ADP-ribosylation is involved in diverse biological processes, it has remained a challenge to identify ADP-ribose acceptor sites. Here, we present an experimental workflow for sensitive and unb...
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Article
Open AccessA continuous sirtuin activity assay without any coupling to enzymatic or chemical reactions
Sirtuins are NAD+ dependent lysine deacylases involved in many regulatory processes such as control of metabolic pathways, DNA repair and stress response. Modulators of sirtuin activity are required as tools for ...
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Article
Open AccessARTD1 regulates osteoclastogenesis and bone homeostasis by dampening NF-κB-dependent transcription of IL-1β
While ADP-ribosyltransferase diphtheria toxin-like 1 (ARTD1, formerly PARP1) and its enzymatic activity have been shown to be important for reprogramming and differentiation of cells, such as during adipogenes...
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Article
Open AccessHyaluronic acid fragments enhance the inflammatory and catabolic response in human intervertebral disc cells through modulation of toll-like receptor 2 signalling pathways
Intervertebral disc (IVD) degeneration is characterized by extracellular matrix breakdown and is considered to be a primary cause of discogenic back pain. Although increases in pro-inflammatory cytokine levels...
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Article
Macrodomain-containing proteins are new mono-ADP-ribosylhydrolases
ADP-ribosylation is an important post-translational protein modification, yet enzymes capable of removing the protein-proximal ADP-ribose were unknown. Human macrodomain proteins MacroD1, D2 and C6orf130 are n...
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Article
Open AccessCrosstalk between SET7/9-dependent methylation and ARTD1-mediated ADP-ribosylation of histone H1.4
Different histone post-translational modifications (PTMs) fine-tune and integrate different cellular signaling pathways at the chromatin level. ADP-ribose modification of histones by cellular ADP-ribosyltransf...