NAD Metabolome Analysis in Human Cells Using 1H NMR Spectroscopy
| العنوان: | NAD Metabolome Analysis in Human Cells Using 1H NMR Spectroscopy |
|---|---|
| المؤلفون: | 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 |
| المصدر: | International Journal of Molecular Sciences, Vol 19, Iss 12, p 3906 (2018) International Journal of Molecular Sciences Volume 19 Issue 12 |
| بيانات النشر: | MDPI AG, 2018. |
| سنة النشر: | 2018 |
| مصطلحات موضوعية: | 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 |
| الوصف: | 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. |
| وصف الملف: | application/pdf |
| اللغة: | English |
| تدمد: | 1422-0067 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::9be2fbda22d65888c80dbc0fd8973625 https://www.mdpi.com/1422-0067/19/12/3906 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....9be2fbda22d65888c80dbc0fd8973625 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 14220067 |
|---|