ELTA: Enzymatic Labeling of Terminal ADP-Ribose

Mol Cell. 2019 Feb 21;73(4):845-856.e5. doi: 10.1016/j.molcel.2018.12.022. Epub 2019 Jan 31.


ADP-ribosylation refers to the addition of one or more ADP-ribose groups onto proteins. The attached ADP-ribose monomers or polymers, commonly known as poly(ADP-ribose) (PAR), modulate the activities of the modified substrates or their binding affinities to other proteins. However, progress in this area is hindered by a lack of tools to investigate this protein modification. Here, we describe a new method named ELTA (enzymatic labeling of terminal ADP-ribose) for labeling free or protein-conjugated ADP-ribose monomers and polymers at their 2'-OH termini using the enzyme OAS1 and dATP. When coupled with various dATP analogs (e.g., radioactive, fluorescent, affinity tags), ELTA can be used to explore PAR biology with techniques routinely used to investigate DNA or RNA function. We demonstrate that ELTA enables the biophysical measurements of protein binding to PAR of a defined length, detection of PAR length from proteins and cells, and enrichment of sub-femtomole amounts of ADP-ribosylated peptides from cell lysates.

Keywords: ADP-ribose; ADP-ribosylated protein; ADP-ribosylation; ADP-ribosyltransferase; enzymatic labeling; mono(ADP-ribosyl)ated protein; oligoadenylate synthetase; poly(ADP-ribose); poly(ADP-ribose) polymerase; poly(ADP-ribosyl)ated protein.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 2',5'-Oligoadenylate Synthetase / genetics
  • 2',5'-Oligoadenylate Synthetase / metabolism*
  • ADP-Ribosylation*
  • Adenosine Diphosphate Ribose / metabolism*
  • Animals
  • Deoxyadenine Nucleotides / metabolism*
  • HeLa Cells
  • Humans
  • Poly(ADP-ribose) Polymerases / metabolism*
  • Protein Binding
  • Protein Domains
  • Sf9 Cells
  • Ubiquitin-Protein Ligases / genetics
  • Ubiquitin-Protein Ligases / metabolism*


  • Deoxyadenine Nucleotides
  • Adenosine Diphosphate Ribose
  • 2'-deoxy-5'-adenosine monophosphate
  • RNF146 protein, human
  • Ubiquitin-Protein Ligases
  • Poly(ADP-ribose) Polymerases
  • OAS1 protein, human
  • 2',5'-Oligoadenylate Synthetase
  • 2'-deoxyadenosine triphosphate