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- W4308790721 endingPage "e202201455" @default.
- W4308790721 startingPage "e202201455" @default.
- W4308790721 abstract "The modification of substrates with ADP-ribose (ADPr) is important in, for example, antiviral immunity and cancer. Recently, several reagents were developed to detect ADP-ribosylation; however, it is unknown whether they recognise ADPr, specific amino acid-ADPr linkages, or ADPr with the surrounding protein backbone. We first optimised methods to prepare extracts containing ADPr-proteins and observe that depending on the amino acid modified, the modification is heatlabile. We tested the reactivity of available reagents with diverse ADP-ribosylated protein and RNA substrates and observed that not all reagents are equally suited for all substrates. Next, we determined cross-reactivity with adenylylated RNA, AMPylated proteins, and metabolites, including NADH, which are detected by some reagents. Lastly, we analysed ADP-ribosylation using confocal microscopy, where depending on the fixation method, either mitochondrion, nucleus, or nucleolus is stained. This study allows future work dissecting the function of ADP-ribosylation in cells, both on protein and on RNA substrates, as we optimised sample preparation methods and have defined the reagents suitable for specific methods and substrates." @default.
- W4308790721 created "2022-11-15" @default.
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- W4308790721 date "2022-11-11" @default.
- W4308790721 modified "2023-09-26" @default.
- W4308790721 title "Protein and RNA ADP-ribosylation detection is influenced by sample preparation and reagents used" @default.
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- W4308790721 doi "https://doi.org/10.26508/lsa.202201455" @default.
- W4308790721 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/36368907" @default.
- W4308790721 hasPublicationYear "2022" @default.
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