TY - JOUR
T1 - Chemical Proteomics Approach for Profiling the NAD Interactome
AU - Šileikytė, Justina
AU - Sundalam, Sunil
AU - David, Larry L.
AU - Cohen, Michael S.
N1 - Funding Information:
We thank Ashok Reddy and John Klimek from the OHSU Proteomics core for assistance with LC–MS/MS analysis. We thank Chadwick Smith and John Koberstein from the OHSU Vollum Institute for advice on differential scanning fluorimetry experiments. We thank Ivan R. Siordia and Daniel Sanderson for protein purification. Mass spectrometric analysis was performed by the OHSU Proteomics Shared Resource with partial support from NIH core grants P30EY010572 & P30CA069533. This work was funded by the NIH (NIH 2R01NS088629) and Fondation Leducq (16CVD04).
Publisher Copyright:
© 2021 American Chemical Society. All rights reserved.
PY - 2021/5/12
Y1 - 2021/5/12
N2 - 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.
AB - 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.
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U2 - 10.1021/jacs.1c01302
DO - 10.1021/jacs.1c01302
M3 - Article
C2 - 33914500
AN - SCOPUS:85106454387
SN - 0002-7863
VL - 143
SP - 6787
EP - 6791
JO - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
IS - 18
ER -