An HPF1/PARP1-Based Chemical Biology Strategy for Exploring ADP-Ribosylation

Cell. 2020 Nov 12;183(4):1086-1102.e23. doi: 10.1016/j.cell.2020.09.055.


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 mono-ADPr. 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.

Keywords: ADP-ribosylation; DNA damage; HFP1; MARylation; PARP1; antibodies; chemical biology; histones; mono-ADP-ribosylation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADP-Ribosylation* / drug effects
  • Amino Acid Sequence
  • Antibodies / metabolism
  • Benzimidazoles / pharmacology
  • Carrier Proteins / metabolism*
  • Cell Line, Tumor
  • Cell Surface Display Techniques
  • DNA Damage
  • Glycoside Hydrolases / metabolism
  • Histones / metabolism
  • Humans
  • Nuclear Proteins / metabolism*
  • Phosphates / metabolism
  • Phosphoric Monoester Hydrolases / metabolism
  • Phthalazines / pharmacology
  • Piperazines / pharmacology
  • Poly (ADP-Ribose) Polymerase-1 / chemistry
  • Poly (ADP-Ribose) Polymerase-1 / metabolism*
  • Recombinant Proteins / metabolism
  • Serine / metabolism
  • Tyrosine / metabolism


  • Antibodies
  • Benzimidazoles
  • Carrier Proteins
  • HPF1 protein, human
  • Histones
  • Nuclear Proteins
  • Phosphates
  • Phthalazines
  • Piperazines
  • Recombinant Proteins
  • veliparib
  • Tyrosine
  • Serine
  • PARP1 protein, human
  • Poly (ADP-Ribose) Polymerase-1
  • Phosphoric Monoester Hydrolases
  • Glycoside Hydrolases
  • ADPRS protein, human
  • olaparib