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Literature summary extracted from

  • Tempel, W.; Rabeh, W.M.; Bogan, K.L.; Belenky, P.; Wojcik, M.; Seidle, H.F.; Nedyalkova, L.; Yang, T.; Sauve, A.A.; Park, H.W.; Brenner, C.
    Nicotinamide riboside kinase structures reveal new pathways to NAD+ (2007), PLoS Biol., 5, e263.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.173 His-tagged wild-type Nrk1 expression in Escherichia coli, expression of wild-type and mutant Nrk1 from the GAL1 promoter on a LEU2 plasmid in Saccharomyces cerevisiae strain BY278 Homo sapiens
2.7.1.173 His-tagged wild-type Nrk2 expression in Escherichia coli, expression of wild-type and mutant Nrk2 from the GAL1 promoter on a LEU2 plasmid in Saccharomyces cerevisiae strain BY278 Homo sapiens

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.7.1.173 Nrk1 bound to nucleoside and nucleotide substrates and products, i.e. nicotinamide mononucleotide, ADP, tiazofurin, beta-D-ribosylnicotinate with AppNHp, and beta-D-ribosylnicotinate alone, addition of 10 mM ligand and 20 mM MgCl2, overview. Usage of different crystallization solutions depending on the ligand, containing PEG 2000 mono-methylether or PEG 3350 or PEG 4000 at 15-35%, at pH 5.0-8.0, with 0.2 M NaH2PO4, and different buffers, X-ray diffraction structure determination and analysis at 1.32-1.95 A resolution, molecular replacement Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
2.7.1.173 D35A site-directed mutagenesis of Nrk2 Homo sapiens
2.7.1.173 D36A site-directed mutagenesis of Nrk1 Homo sapiens
2.7.1.173 E100A site-directed mutagenesis of Nrk2, mutant activity in ATP-dependent phosphorylation is below the level of detection Homo sapiens
2.7.1.173 E174R site-directed mutagenesis of Nrk2 Homo sapiens
2.7.1.173 E98A site-directed mutagenesis of Nrk1, mutant activity in ATP-dependent phosphorylation is below the level of detection Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.1.173 0.051
-
 beta-D-ribosylnicotinamide Nrk1, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 0.063
-
 beta-D-ribosylnicotinamide Nrk2, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 0.068
-
 beta-D-ribosylnicotinate Nrk1, with GTP, pH 7.5, 37°C Homo sapiens
2.7.1.173 0.088
-
 beta-D-ribosylnicotinate Nrk1, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 0.11
-
 tiazofurin Nrk2, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 0.19
-
 beta-D-ribosylnicotinate Nrk2, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 0.27
-
 tiazofurin Nrk1, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 1.3
-
 uridine Nrk2, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 15
-
 cytidine Nrk2, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 17
-
 uridine Nrk1, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 30
-
 cytidine Nrk1, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 30
-
 beta-D-ribosylnicotinate Nrk2, with GTP, pH 7.5, 37°C Homo sapiens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.1.173 Mg2+ required Homo sapiens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.1.173 ATP + beta-D-ribosylnicotinate Homo sapiens nicotinic acid riboside is a specific substrate of human Nrk enzymes ADP + nicotinate beta-D-ribonucleotide
-
 ?
2.7.1.173 additional information Homo sapiens Asp35 and Glu100 in Nrk2 are essential residues for function in vivo ?
-
 ?
2.7.1.173 additional information Homo sapiens Asp36 and Glu98 in Nrk1 are essential residues for function in vivo ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.173 Homo sapiens Q9NPI5 Nrk2; Nrk2
-
2.7.1.173 Homo sapiens Q9NWW6 Nrk1; Nrk1
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.173 recombinant Nrk1 and Nrk2 from Escherichia coli by metal chelate affinity chromatography Homo sapiens
2.7.1.173 recombinant Nrk2 from Escherichia coli by metal chelate affinity chromatography Homo sapiens

Source Tissue

EC Number Source Tissue Comment Organism Textmining

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.173 ATP + beta-D-ribosylnicotinamide beta-D-ribosylnicotinamide is an NAD+ precursor utilized via the Nrk and Preiss-Hander pathways Homo sapiens ADP + nicotinamide beta-D-ribonucleotide
-
 ?
2.7.1.173 ATP + beta-D-ribosylnicotinate nicotinic acid riboside is a specific substrate of human Nrk enzymes Homo sapiens ADP + nicotinate beta-D-ribonucleotide
-
 ?
2.7.1.173 ATP + cytidine
-
 Homo sapiens ADP + CMP
-
 ?
2.7.1.173 ATP + cytidine low activity with Nrk1 Homo sapiens ADP + CMP
-
 ?
2.7.1.173 ATP + nicotinamide mononucleotide
-
 Homo sapiens ADP + nicotinamide mononucleotide phosphate
-
 ?
2.7.1.173 ATP + tiazofurin
-
 Homo sapiens ADP + ?
-
 ?
2.7.1.173 ATP + tiazofurin a relatively good Nrk1 substrate Homo sapiens ADP + ?
-
 ?
2.7.1.173 ATP + uridine
-
 Homo sapiens ADP + UMP
-
 ?
2.7.1.173 ATP + uridine low activity with Nrk1 Homo sapiens ADP + UMP
-
 ?
2.7.1.173 GTP + beta-D-ribosylnicotinate nicotinic acid riboside is a specific substrate of human Nrk enzymes Homo sapiens GDP + nicotinate beta-D-ribonucleotide
-
 ?
2.7.1.173 additional information Asp35 and Glu100 in Nrk2 are essential residues for function in vivo Homo sapiens ?
-
 ?
2.7.1.173 additional information Asp36 and Glu98 in Nrk1 are essential residues for function in vivo Homo sapiens ?
-
 ?
2.7.1.173 additional information substrate specificity of recombinant Nrk1 and Nrk2, overview. Classification of Nrk1 as an nicotinic acid riboside and tiazofurin:ATP or GTP kinase, Nrk1 displays a 340fold preference for nicotinic acid riboside over cytidine in the KM term and a 500fold preference over either cytidine or uridine in the kcat/KM term. Poor cytidine monophosphate-forming activity of the enzyme. Structural basis of substrate specificity, overview. Base recognition in the Nrk1 nucleoside-binding site excludes uridine but supports beta-D-ribosylnicotinamide phosphorylation Homo sapiens ?
-
 ?
2.7.1.173 additional information substrate specificity of recombinant Nrk2, overview. Classification of Nrk2 as an ATP-specific nicotinic acid riboside, tiazofurin, and uridine kinase. Poor cytidine monophosphate-forming activity of the enzyme. Structural basis of substrate specificity, overview Homo sapiens ?
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.1.173 More the ADP-binding site includes P-loop sequence Gly-Val-Thr-Asn-Ser-Gly-Lys-Thr, residues 10-17 Homo sapiens

Synonyms

EC Number Synonyms Comment Organism
2.7.1.173 nicotinamide riboside kinase
-
 Homo sapiens
2.7.1.173 NRK1
-
 Homo sapiens
2.7.1.173 Nrk2
-
 Homo sapiens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.1.173 37
-
 assay at Homo sapiens

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.1.173 0.21
-
 uridine Nrk1, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 0.21
-
 beta-D-ribosylnicotinamide Nrk1, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 0.34
-
 beta-D-ribosylnicotinate Nrk1, with GTP, pH 7.5, 37°C Homo sapiens
2.7.1.173 0.34
-
 beta-D-ribosylnicotinamide Nrk2, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 0.35
-
 tiazofurin Nrk1, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 0.48
-
 cytidine Nrk1, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 0.49
-
 tiazofurin Nrk2, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 0.6
-
 beta-D-ribosylnicotinate Nrk1, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 0.75
-
 beta-D-ribosylnicotinate Nrk2, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 0.82
-
 cytidine Nrk2, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 1.1
-
 uridine Nrk2, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 1.7
-
 beta-D-ribosylnicotinate Nrk2, with GTP, pH 7.5, 37°C Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.1.173 7.5
-
 assay at Homo sapiens

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.1.173 ATP active with Nrk1 Homo sapiens
2.7.1.173 ATP specific for Nrk2 Homo sapiens
2.7.1.173 GTP active with Nrk1 Homo sapiens
2.7.1.173 additional information GTP is not with Nrk2 Homo sapiens

General Information

EC Number General Information Comment Organism
2.7.1.173 evolution the enzyme is structurally similar to Rossmann fold metabolite kinases Homo sapiens
2.7.1.173 metabolism the enzyme is involved in the eukaryotic nicotinamide riboside kinase, Nrk, pathway, which is induced in response to nerve damage and promotes replicative life span in yeast, converts nicotinamide riboside to NAD+ by phosphorylation and adenylylation, overview. Nicotinic acid riboside is utilized in vivo by Urh1, Pnp1, and Preiss-Handler salvage Homo sapiens

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.7.1.173 0.012
-
 uridine Nrk1, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 0.016
-
 cytidine Nrk1, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 0.055
-
 cytidine Nrk2, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 0.057
-
 beta-D-ribosylnicotinate Nrk2, with GTP, pH 7.5, 37°C Homo sapiens
2.7.1.173 0.85
-
 uridine Nrk2, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 1.3
-
 tiazofurin Nrk1, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 3.9
-
 beta-D-ribosylnicotinate Nrk2, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 4.1
-
 beta-D-ribosylnicotinamide Nrk1, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 4.5
-
 tiazofurin Nrk2, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 5
-
 beta-D-ribosylnicotinate Nrk1, with GTP, pH 7.5, 37°C Homo sapiens
2.7.1.173 5.4
-
 beta-D-ribosylnicotinamide Nrk2, with ATP, pH 7.5, 37°C Homo sapiens
2.7.1.173 6.8
-
 beta-D-ribosylnicotinate Nrk1, with ATP, pH 7.5, 37°C Homo sapiens