NAD Metabolome Analysis in Human Cells Using 1H NMR Spectroscopy

Autor: Veronika Kulikova, Kirill Nerinovski, L. V. Solovjeva, Mikhail Khodorkovskiy, Mathias Ziegler, Maria Svetlova, Andrey Nikiforov, K. V. Shabalin, Alexander Yakimov, Marie E. Migaud, Stepan Gambaryan, Richard Cunningham
Jazyk: angličtina
Rok vydání: 2018
Předmět:
Blood Platelets
Niacinamide
0301 basic medicine
NAD metabolome
Erythrocytes
Proton Magnetic Resonance Spectroscopy
Cell Culture Techniques
Nicotinamide adenine dinucleotide
Niacin
Redox
Article
Catalysis
Cofactor
Inorganic Chemistry
lcsh:Chemistry
03 medical and health sciences
chemistry.chemical_compound
0302 clinical medicine
NMR spectroscopy
Humans
Metabolomics
Physical and Theoretical Chemistry
Molecular Biology
lcsh:QH301-705.5
Spectroscopy
Nicotinamide
biology
human cells
Organic Chemistry
General Medicine
Riboside
NAD
Computer Science Applications
Vitamin B3
HEK293 Cells
030104 developmental biology
chemistry
Biochemistry
lcsh:Biology (General)
lcsh:QD1-999
030220 oncology & carcinogenesis
Nicotinamide riboside
biology.protein
NAD+ kinase
Metabolic Networks and Pathways
NADP
Protein deacetylation
Zdroj: International Journal of Molecular Sciences, Vol 19, Iss 12, p 3906 (2018)
International Journal of Molecular Sciences
Volume 19
Issue 12
ISSN: 1422-0067
Popis: Nicotinamide adenine dinucleotide (NAD) and its phosphorylated form, NADP, are the major coenzymes of redox reactions in central metabolic pathways. Nicotinamide adenine dinucleotide is also used to generate second messengers, such as cyclic ADP-ribose, and serves as substrate for protein modifications including ADP-ribosylation and protein deacetylation by sirtuins. The regulation of these metabolic and signaling processes depends on NAD availability. Generally, human cells accomplish their NAD supply through biosynthesis using different forms of vitamin B3: Nicotinamide (Nam) and nicotinic acid as well as nicotinamide riboside (NR) and nicotinic acid riboside (NAR). These precursors are converted to the corresponding mononucleotides NMN and NAMN, which are adenylylated to the dinucleotides NAD and NAAD, respectively. Here, we have developed an NMR-based experimental approach to detect and quantify NAD(P) and its biosynthetic intermediates in human cell extracts. Using this method, we have determined NAD, NADP, NMN and Nam pools in HEK293 cells cultivated in standard culture medium containing Nam as the only NAD precursor. When cells were grown in the additional presence of both NAR and NR, intracellular pools of deamidated NAD intermediates (NAR, NAMN and NAAD) were also detectable. We have also tested this method to quantify NAD+ in human platelets and erythrocytes. Our results demonstrate that 1H NMR spectroscopy provides a powerful method for the assessment of the cellular NAD metabolome.
Databáze: OpenAIRE
Externí odkaz:
Nepřihlášeným uživatelům se plný text nezobrazuje