Chemical Proteomics Approach for Profiling the NAD Interactome

Area of Science:

• Biochemistry
• Chemical Biology
• Proteomics

Background:

• Nicotinamide adenine dinucleotide (NAD+) is crucial for metabolism and serves as a substrate for enzymes like poly-ADP-ribose polymerases (PARPs).
• The full scope of NAD+ interactions, known as the NAD interactome, is likely underestimated due to a lack of effective chemical tools for its exploration.

Purpose of the Study:

• To design, synthesize, and validate novel clickable photoaffinity labeling (PAL) probes for profiling NAD+-binding proteins.
• To identify new components of the NAD interactome using these chemical tools.

Main Methods:

• Development of clickable PAL probes, specifically 2- and 6-ad-BAD, based on NAD+.
• Application of the probes in chemical proteomics experiments to identify NAD+-binding proteins.
• UV-dependent labeling of proteins by the 2-ad-BAD probe.

Main Results:

• The 2-ad-BAD probe demonstrated efficient, UV-dependent labeling of PARPs.
• Proteomics studies using both 2- and 6-ad-BAD identified previously known and novel NAD+/NADH-binding proteins.
• Validation of the probes' utility in interrogating the NAD interactome.

Conclusions:

• The developed clickable PAL probes, 2- and 6-ad-BAD, are effective tools for investigating the NAD interactome.
• These probes facilitate the discovery of new NAD+-binding proteins, advancing the understanding of NAD+ roles in cellular processes.