Affinity-based protein profiling-driven discovery of myricanol as a Nampt activator

  • Bioorg Chem. 2023 Apr:133:106435. doi: 10.1016/j.bioorg.2023.106435.
Peng Lyu  1 ,  Shengrong Li  2 ,  Ying Han  3 ,  Shengnan Shen  1 ,  Zheling Feng  1 ,  Piliang Hao  4 ,  Zhengqiu Li  5 ,  Ligen Lin  6
Affiliations
  • 1. State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Taipa, Macau 999078, China.
  • 2. International Cooperative Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug Development (MOE), MOE Key Laboratory of Tumor Molecular Biology, School of Pharmacy, Jinan University, Guangzhou 510632, China.
  • 3. School of Life Science and Technology, ShanghaiTech University, 393 Middle Huaxia Road, Shanghai 201210, China.
  • 4. School of Life Science and Technology, ShanghaiTech University, 393 Middle Huaxia Road, Shanghai 201210, China. Electronic address: [email protected].
  • 5. International Cooperative Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug Development (MOE), MOE Key Laboratory of Tumor Molecular Biology, School of Pharmacy, Jinan University, Guangzhou 510632, China. Electronic address: [email protected].
  • 6. State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Taipa, Macau 999078, China. Electronic address: [email protected].
PMID: 36841049 DOI: 10.1016/j.bioorg.2023.106435
Abstract

Herein, we synthesized an affinity-based probe of myricanol (pMY) with a photo-affinity cross-linker to initiate a bioconjugation reaction, which was applied for target identification in live C2C12 myotubes. Pull-down of biotinylated pMY coupled with mass spectroscopy and Western blotting revealed that pMY can bind with nicotinamide phosphoribosyltransferase (NAMPT), a rate-limiting enzyme in the nicotinamide adenine dinucleotide salvage pathway. Cellular thermal shift assay, drug affinity responsive target stability assay and recombinant protein labeling further validated the direct interaction between myricanol and NAMPT. Myricanol did not affect the protein expression of NAMPT, but enhanced its activity. Knock-down of NAMPT totally abolished the promoting effect of myricanol on insulin-stimulated glucose uptake in C2C12 myotubes. Taken together, myricanol sensitizes Insulin action in myotubes through binding with and activating NAMPT.

Keywords
Activity-based protein profiling; Myotubes; Myricanol; Nampt; Photo-affinity probe.
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