Chemical Proteomics Approach for Profiling the NAD Interactome

J Am Chem Soc. 2021 May 12;143(18):6787-6791. doi: 10.1021/jacs.1c01302. Epub 2021 Apr 29.

Abstract

Nicotinamide adenine dinucleotide (NAD+) is a multifunctional molecule. Beyond redox metabolism, NAD+ has an equally important function as a substrate for post-translational modification enzymes, the largest family being the poly-ADP-ribose polymerases (PARPs, 17 family members in humans). The recent surprising discoveries of noncanonical NAD (NAD+/NADH)-binding proteins suggests that the NAD interactome is likely larger than previously thought; yet, broadly useful chemical tools for profiling and discovering NAD-binding proteins do not exist. Here, we describe the design, synthesis, and validation of clickable, photoaffinity labeling (PAL) probes, 2- and 6-ad-BAD, for interrogating the NAD interactome. We found that 2-ad-BAD efficiently labels PARPs in a UV-dependent manner. Chemical proteomics experiments with 2- and 6-ad-BAD identified known and unknown NAD+/NADH-binding proteins. Together, our study shows the utility of 2- and 6-ad-BAD as clickable PAL NAD probes.

Publication types

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

MeSH terms

  • Adenine Nucleotides / chemistry*
  • Benzamides / chemistry*
  • Carrier Proteins / chemistry*
  • Humans
  • NAD / chemistry*
  • Proteomics*

Substances

  • Adenine Nucleotides
  • Benzamides
  • Carrier Proteins
  • NAD
  • benzamide adenine nucleotide