Equilibrative Nucleoside Transporters Mediate the Import of Nicotinamide Riboside and Nicotinic Acid Riboside into Human Cells.

Publisher: MDPI Country of Publication: Switzerland NLM ID: 101092791 Publication Model: Electronic Cited Medium: Internet ISSN: 1422-0067 (Electronic) Linking ISSN: 14220067 NLM ISO Abbreviation: Int J Mol Sci Subsets: MEDLINE

Original Publication: Basel, Switzerland : MDPI, [2000-

Equilibrative Nucleoside Transport Proteins/*metabolism ; Membrane Transport Proteins/*metabolism ; Niacinamide/*analogs & derivatives ; Pyridinium Compounds/*metabolism ; Ribonucleosides/*metabolism; Aging/metabolism ; Cytosol/metabolism ; Equilibrative Nucleoside Transport Proteins/genetics ; HEK293 Cells ; Humans ; Magnetic Resonance Spectroscopy ; Membrane Transport Proteins/analysis ; Membrane Transport Proteins/genetics ; Metabolomics ; NAD/analysis ; NAD/metabolism ; Niacinamide/analysis ; Niacinamide/metabolism ; Nicotinamide Mononucleotide/metabolism ; Phosphorylation/physiology ; Phosphotransferases (Alcohol Group Acceptor)/metabolism ; Pyridinium Compounds/analysis ; Recombinant Proteins/genetics ; Recombinant Proteins/metabolism ; Ribonucleosides/analysis

Nicotinamide riboside (NR), a new form of vitamin B3, is an effective precursor of nicotinamide adenine dinucleotide (NAD ⁺ ) in human and animal cells. The introduction of NR into the body effectively increases the level of intracellular NAD ⁺ and thereby restores physiological functions that are weakened or lost in experimental models of aging and various pathologies. Despite the active use of NR in applied biomedicine, the mechanism of its transport into mammalian cells is currently not understood. In this study, we used overexpression of proteins in HEK293 cells, and metabolite detection by NMR, to show that extracellular NR can be imported into cells by members of the equilibrative nucleoside transporter (ENT) family ENT1, ENT2, and ENT4. After being imported into cells, NR is readily metabolized resulting in Nam generation. Moreover, the same ENT-dependent mechanism can be used to import the deamidated form of NR, nicotinic acid riboside (NAR). However, NAR uptake into HEK293 cells required the stimulation of its active utilization in the cytosol such as phosphorylation by NR kinase. On the other hand, we did not detect any NR uptake mediated by the concentrative nucleoside transporters (CNT) CNT1, CNT2, or CNT3, while overexpression of CNT3, but not CNT1 or CNT2, moderately stimulated NAR utilization by HEK293 cells.

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19-34-60039 Russian Foundation for Basic Research; 250395 Norges Forskningsråd

Keywords: NAD+ metabolism; concentrative nucleoside transporters; equilibrative nucleoside transporters; human cells; nicotinamide riboside; nicotinic acid riboside

0 (Equilibrative Nucleoside Transport Proteins)
0 (Membrane Transport Proteins)
0 (Pyridinium Compounds)
0 (Recombinant Proteins)
0 (Ribonucleosides)
0 (cif nucleoside transporter)
0I8H2M0L7N (nicotinamide-beta-riboside)
0U46U6E8UK (NAD)
1094-61-7 (Nicotinamide Mononucleotide)
17720-18-2 (nicotinic acid ribonucleoside)
25X51I8RD4 (Niacinamide)
EC 2.7.1.- (Phosphotransferases (Alcohol Group Acceptor))
EC 2.7.1.22 (NMRK1 protein, human)

Date Created: 20210212 Date Completed: 20210907 Latest Revision: 20210907

20231215

PMC7866510

10.3390/ijms22031391

33573263