A ribose-functionalized NAD+ with unexpected high activity and selectivity for protein poly-ADP-ribosylation.
Nature communications
confidence
Key findings
3'-azido NAD+ analogue synthesized with high efficiency; exhibits high activity and specificity for PARP1/2-mediated protein PARylation and enhanced labeling.
View source on PubMed (PMID 31519936) ↗
- Sample size
- Not reported
- Population
- In vitro (enzymatic synthesis and PARylation assays); cellular contexts for labeling
- Dosing
- Not reported
- Duration
- Not reported
- Route
- Not applicable (in vitro)
- Blinding
- not_reported
- Controls
- none
- Drug class
- coenzyme
Full abstract
Nicotinamide adenine dinucleotide (NAD+)-dependent ADP-ribosylation plays important roles in physiology and pathophysiology. It has been challenging to study this key type of enzymatic post-translational modification in particular for protein poly-ADP-ribosylation (PARylation). Here we explore chemical and chemoenzymatic synthesis of NAD+ analogues with ribose functionalized by terminal alkyne and azido groups. Our results demonstrate that azido substitution at 3'-OH of nicotinamide riboside enables enzymatic synthesis of an NAD+ analogue with high efficiency and yields. Notably, the generated 3'-azido NAD+ exhibits unexpected high activity and specificity for protein PARylation catalyzed by human poly-ADP-ribose polymerase 1 (PARP1) and PARP2. And its derived poly-ADP-ribose polymers show increased resistance to human poly(ADP-ribose) glycohydrolase-mediated degradation. These unique properties lead to enhanced labeling of protein PARylation by 3'-azido NAD+ in the cellular contexts and facilitate direct visualization and labeling of mitochondrial protein PARylation. The 3'-azido NAD+ provides an important tool for studying cellular PARylation.