{"_id":"HEkkHcoaZSnmwunnB","bibbaseid":"anonymous-nadmetabolomeanalysisinhumancellsusing1hnmrspectroscopy-2018","authorIDs":[],"bibdata":{"bibtype":"article","type":"article","author":[{"propositions":[],"lastnames":["Shabalin"],"firstnames":["K."],"suffixes":[]},{"propositions":[],"lastnames":["Nerinovski"],"firstnames":["K."],"suffixes":[]},{"propositions":[],"lastnames":["Yakimov"],"firstnames":["A."],"suffixes":[]},{"propositions":[],"lastnames":["Kulikova"],"firstnames":["V."],"suffixes":[]},{"propositions":[],"lastnames":["Svetlova"],"firstnames":["M."],"suffixes":[]},{"propositions":[],"lastnames":["Solovjeva"],"firstnames":["L."],"suffixes":[]},{"propositions":[],"lastnames":["Khodorkovskiy"],"firstnames":["M."],"suffixes":[]},{"propositions":[],"lastnames":["Gambaryan"],"firstnames":["S."],"suffixes":[]},{"propositions":[],"lastnames":["Cunningham"],"firstnames":["R."],"suffixes":[]},{"propositions":[],"lastnames":["Migaud"],"firstnames":["M.E."],"suffixes":[]},{"propositions":[],"lastnames":["Ziegler"],"firstnames":["M."],"suffixes":[]},{"propositions":[],"lastnames":["Nikiforov"],"firstnames":["A."],"suffixes":[]}],"title":"NAD metabolome analysis in human cells using 1 H NMR spectroscopy","journal":"International Journal of Molecular Sciences","year":"2018","volume":"19","number":"12","doi":"10.3390/ijms19123906","art_number":"3906","note":"cited By 0","url":"https://www.scopus.com/inward/record.uri?eid=2-s2.0-85058324244&doi=10.3390%2fijms19123906&partnerID=40&md5=fea565a236f1148eb16893f161907a87","affiliation":"Institute of Cytology, Russian Academy of Sciences, St. Petersburg, 194064, Russian Federation; Petersburg Nuclear Physics Institute, NRC Kurchatov Institute, Gatchina, 188300, Russian Federation; Department of Nuclear Physics Research Methods, St. Petersburg State University, St. Petersburg, 199034, Russian Federation; Peter the Great St. Petersburg Polytechnic University, St. Petersburg, 195251, Russian Federation; Sechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, St. Petersburg, 194223, Russian Federation; Mitchell Cancer Institute, University of South Alabama, Mobile, AL 36604, United States; Department of Biomedicine, University of Bergen, Bergen, 5020, Norway","abstract":"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 1 H NMR spectroscopy provides a powerful method for the assessment of the cellular NAD metabolome. © 2018 by the authors. Licensee MDPI, Basel, Switzerland.","author_keywords":"Human cells; NAD metabolome; NMR spectroscopy; Vitamin B3","funding_details":"Russian Science Foundation16-14-10240","key":"Shabalin2018","id":"Shabalin2018","bibbaseid":"anonymous-nadmetabolomeanalysisinhumancellsusing1hnmrspectroscopy-2018","role":"author","urls":{"Paper":"https://www.scopus.com/inward/record.uri?eid=2-s2.0-85058324244&doi=10.3390%2fijms19123906&partnerID=40&md5=fea565a236f1148eb16893f161907a87"},"metadata":{"authorlinks":{}},"downloads":0},"bibtype":"article","biburl":"https://bio.pnpi.nrcki.ru/wp-content/uploads/2019/12/lbm_2019_10.txt","creationDate":"2019-11-05T13:18:15.228Z","downloads":0,"keywords":[],"search_terms":["nad","metabolome","analysis","human","cells","using","nmr","spectroscopy"],"title":"NAD metabolome analysis in human cells using 1 H NMR spectroscopy","year":2018,"dataSources":["YYLjupmCazkNqWNEJ"]}