ELTA: Enzymatic Labeling of Terminal ADP-Ribose.
Mol Cell
; 73(4): 845-856.e5, 2019 02 21.
Article
in En
| MEDLINE
| ID: mdl-30712989
ABSTRACT
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.
Key words
Full text:
1
Database:
MEDLINE
Main subject:
2',5'-Oligoadenylate Synthetase
/
Adenosine Diphosphate Ribose
/
Poly(ADP-ribose) Polymerases
/
Ubiquitin-Protein Ligases
/
Deoxyadenine Nucleotides
/
ADP-Ribosylation
Limits:
Animals
/
Humans
Language:
En
Journal:
Mol Cell
Journal subject:
BIOLOGIA MOLECULAR
Year:
2019
Type:
Article
Affiliation country:
United States