期刊
CELL
卷 183, 期 4, 页码 1086-+出版社
CELL PRESS
DOI: 10.1016/j.cell.2020.09.055
关键词
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资金
- Germany's Excellence Strategy (CECAD) [EXC 2030-390661388]
- Max Planck Society, Germany
- EU's Horizon 2020 program (Marie Sklodowska-Curie) [657501]
- EMBO Young Investigator Programme
- Cologne Graduate School of Ageing Research
- Deutsche Krebshilfe, Germany, through a Mildred Scheel Nachwuchszentrum [70113307]
- Marie Curie Actions (MSCA) [657501] Funding Source: Marie Curie Actions (MSCA)
Strategies for installing authentic ADP-ribosylation (ADPr) at desired positions are fundamental for creating the tools needed to explore this elusive post-translational modification (PTM) in essential cellular processes. Here, we describe a phospho-guided chemoenzymatic approach based on the Ser-ADPr writer complex for rapid, scalable preparation of a panel of pure, precisely modified peptides. Integrating this methodology with phage display technology, we have developed site-specific as well as broad-specificity antibodies to monoADPr. These recombinant antibodies have been selected and characterized using multiple ADP-ribosylated peptides and tested by immunoblotting and immunofluorescence for their ability to detect physiological ADPr events. Mono-ADPr proteomics and poly-to-mono comparisons at the modification site level have revealed the prevalence of mono-ADPr upon DNA damage and illustrated its dependence on PARG and ARH3. These and future tools created on our versatile chemical biology-recombinant antibody platform have broad potential to elucidate ADPr signaling pathways in health and disease.
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