Quantitative dynamics of intracellular NMN by genetically encoded biosensor

Nicotinamide mononucleotide (NMN) is the direct precursor and a major booster of NAD+ with increasing applications in NAD+- and aging-related pathologies. However, measuring live cell NMN dynamics was not possible, leaving key questions in NMN uptake and intracellular regulation unanswered. Here we...

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Bibliographic Details
Published in:Biosensors & bioelectronics 2025-01, Vol.267, p.116842, Article 116842
Main Authors: Chen, Liuqing, Wang, Pei, Huang, Guan, Cheng, Wenxiang, Liu, Kaijing, Yu, Qiuliyang
Format: Article
Language:English
Subjects:
Biosensing Techniques - methods
Biosensor
HEK293 Cells
Humans
Metabolism
Mitochondria - genetics
Mitochondria - metabolism
NAD
NAD - analysis
NAD - metabolism
Nicotinamide Mononucleotide - chemistry
Nicotinamide Mononucleotide - metabolism
NMN
Subcellular
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Summary:Nicotinamide mononucleotide (NMN) is the direct precursor and a major booster of NAD+ with increasing applications in NAD+- and aging-related pathologies. However, measuring live cell NMN dynamics was not possible, leaving key questions in NMN uptake and intracellular regulation unanswered. Here we developed genetically encoded bioluminescent and fluorescent sensors to quantify subcellular NMN in live cells by engineering specific NMN-responsive protein scaffolds fused to luciferase and fluorescent proteins. The sensor dissected the multimechanistic uptake of exogenous NMN and nicotinamide riboside (NR) in live cells and further measured the NMN levels across different subcellular compartments, as well as the perturbed NMN/NAD+ ratios by external supplements. Moreover, we measured the NMN regulation by NAD(H) hydrolase Nudts and peroxisomal carrier Pxmp2 and identified Slc25a45 as a potential mitochondrial NMN regulator for its unique fingerprint on the local NMN/NAD+ ratio. Collectively, the genetically encoded sensors provide a useful tool for visualizing NMN metabolism.
ISSN:0956-5663
1873-4235
1873-4235
DOI:10.1016/j.bios.2024.116842