A ribose-functionalized NAD + with unexpected high activity and selectivity for protein poly-ADP-ribosylation

Nat Commun. 2019 Sep 13;10(1):4196. doi: 10.1038/s41467-019-12215-4.

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.

Publication types

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

MeSH terms

  • ADP Ribose Transferases / metabolism
  • Chromatography, High Pressure Liquid
  • HeLa Cells
  • Humans
  • Magnetic Resonance Spectroscopy
  • Models, Biological
  • NAD / metabolism*
  • Nicotinamide-Nucleotide Adenylyltransferase / metabolism
  • Phosphotransferases (Alcohol Group Acceptor) / metabolism
  • Poly (ADP-Ribose) Polymerase-1 / metabolism
  • Poly ADP Ribosylation
  • Poly(ADP-ribose) Polymerases / metabolism
  • Proto-Oncogene Proteins / metabolism
  • Sirtuin 2 / metabolism

Substances

  • Proto-Oncogene Proteins
  • NAD
  • ADP Ribose Transferases
  • PARP1 protein, human
  • PARP10 protein, human
  • Poly (ADP-Ribose) Polymerase-1
  • Poly(ADP-ribose) Polymerases
  • Phosphotransferases (Alcohol Group Acceptor)
  • NMRK1 protein, human
  • NMNAT1 protein, human
  • Nicotinamide-Nucleotide Adenylyltransferase
  • SIRT2 protein, human
  • Sirtuin 2