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Literature summary extracted from

  • Hara, N.; Yamada, K.; Shibata, T.; Osago, H.; Hashimoto, T.; Tsuchiya, M.
    Elevation of cellular NAD levels by nicotinic acid and involvement of nicotinic acid phosphoribosyltransferase in human cells (2007), J. Biol. Chem., 282, 24574-24582.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.3.4.21 cDNA fragment (as probe for Northern blot analysis) reverse transcribed from tissue total RNA using sense primer 5’-GTG AGG TGA ATG TCA TTG GC-3’ and antisense primer 5’-ACA GTG CGA CCG GAT ACA CT-3’ Mus musculus
6.3.4.21 expression in Escherichia coli as His6-tagged protein Homo sapiens
6.3.4.21 in pET22b and pET15b for expression in Escherichia coli BL21 (DE3) as fusion protein with C-terminal or N-terminal hexa-His-tag, respectively, in pcDNA3His(6) for expression in eukaryotic cell culture, specific siRNAs against nt829-1634 and nt445-935 for transfection into HEK-293 cells Homo sapiens

Inhibitors

EC Number Inhibitors Comment Organism Structure
6.3.4.21 additional information not inhibited by up to 1 mM NAD+, lack of feedback inhibition by NAD+ Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.3.4.21 0.00022
-
 5-phospho-alpha-D-ribose 1-diphosphate +/-0.07 microM, in presence of 1 mM NAD+, 20 ng enzyme, 40 microM nicotinamide, 0.14-1 microM PRPP, 15 min, 37°C, TLC-based quantification Homo sapiens
6.3.4.21 0.00025
-
 5-phospho-alpha-D-ribose 1-diphosphate +/-0.06 microM, 20 ng enzyme, 40 microM nicotinamide, 0.14-1 microM PRPP, 15 min, 37°C, TLC-based quantification Homo sapiens
6.3.4.21 0.0127
-
 nicotinic acid +/-3.6 microM, in presence of 1 mM NAD+, 80 ng enzyme, 10-70 microM nicotinamide, 0.3 mM PRPP, 30 min, 37°C, TLC-based quantification Homo sapiens
6.3.4.21 0.0155
-
 nicotinic acid +/-3.4 microM, 80 ng enzyme, 10-70 microM nicotinamide, 0.3 mM PRPP, 30 min, 37°C, TLC-based quantification Homo sapiens

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
6.3.4.21 cytosol
-
 Homo sapiens 5829
-
6.3.4.21 cytosol subcellular fractionation, revealed by Western blot and activity measurements Rattus norvegicus 5829
-
6.3.4.21 cytosol when overexpressed in Hep-G2 cells and detected by mouse polyclonal anti-human NAPRT antibody Homo sapiens 5829
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.3.4.21 Mg2+ omission of Mg2+ from the reaction mixture results in a complete loss of activity Homo sapiens
6.3.4.21 Mg2+ omission leads to complete loss of NaMN synthesis activity Homo sapiens

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
6.3.4.21 51000
-
 endogenous NAPRT detected in Western blot using mouse polyclonal anti-human NAPRT antibody Homo sapiens
6.3.4.21 51000
-
 endogenous NAPRT detected in Western blot using mouse polyclonal anti-human NAPRT antibody Rattus norvegicus
6.3.4.21 52000
-
 overexpressed, His-tagged NAPRT in Hep-G2 cells detected in Western blot using mouse polyclonal anti-human NAPRT antibody Homo sapiens
6.3.4.21 54000
-
 purified recombinant NAPRT Homo sapiens
6.3.4.21 55000
-
 2 * 55000, non-denaturing polyacrylamide gel electrophoresis of purified recombinant NAPRT in active state Homo sapiens
6.3.4.21 59800
-
 calculated from the deduced protein sequence Homo sapiens
6.3.4.21 110000
-
 non-denaturing polyacrylamide gel electrophoresis of purified recombinant NAPRT in active state as controlled by applying gel slice to TLC-based activity assay with 50 microM nicotinic acid + 0.3 mM PRPP, 3 h, 37°C Homo sapiens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.3.4.21 nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + ATP + H2O Mus musculus
-
 beta-nicotinate D-ribonucleotide + diphosphate + ADP + phosphate
-
 ?
6.3.4.21 nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + ATP + H2O Homo sapiens
-
 beta-nicotinate D-ribonucleotide + diphosphate + ADP + phosphate
-
 r
6.3.4.21 nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + ATP + H2O Rattus norvegicus
-
 beta-nicotinate D-ribonucleotide + diphosphate + ADP + phosphate
-
 r
6.3.4.21 nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + ATP + H2O Mus musculus
-
 beta-nicotinate D-ribonucleotide + diphosphate + ADP + phosphate
-
 r
6.3.4.21 nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + ATP + H2O Homo sapiens NAPRT is essential for nicotinic acid to increase cellular NAD levels and thus to prevent oxidative stress of the cells beta-nicotinate D-ribonucleotide + diphosphate + ADP + phosphate
-
 ?
6.3.4.21 nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + ATP + H2O Mus musculus BALB/c
-
 beta-nicotinate D-ribonucleotide + diphosphate + ADP + phosphate
-
 r

Organism

EC Number Organism UniProt Comment Textmining
6.3.4.21 Homo sapiens Q6XQN6
-
-
6.3.4.21 Mus musculus
-
-
-
6.3.4.21 Mus musculus Q8CC86 BALB/c
-
6.3.4.21 Mus musculus BALB/c Q8CC86 BALB/c
-
6.3.4.21 Rattus norvegicus Q6XQN1 female Wistar rats
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.3.4.21
-
 Homo sapiens
6.3.4.21 nickel chelate His-bind resin after bacterial expression Homo sapiens

Source Tissue

EC Number Source Tissue Comment Organism Textmining
6.3.4.21 heart
-
 Mus musculus
-
6.3.4.21 heart 1.9 kb mRNA detected by Northern blot analysis, moderate expression Mus musculus
-
6.3.4.21 heart significant endogenous activity Rattus norvegicus
-
6.3.4.21 HEK-293 cell exogenously added nicotinic acid (1-10 microM) leads to increase in basal total NAD++ level (revealed by ESI-MS) in a dose-dependent manner and to NaAD+ accumulation which are reduced by siRNA-based knock-down of endogenous NAPRT, knock-down does not affect basal NAD++ levels (503 +/-104 microM) in absence of exogenous nicotinic acid, exogenously added nicotinic acid decreases oxidative cytotoxicity (by 30-50 microM hydrogen peroxide) through elevated NAD+ levels mediated by NAPRT activity as revealed by siRNA-based knock-down of NAPRT Homo sapiens
-
6.3.4.21 HeLa cell
-
 Homo sapiens
-
6.3.4.21 Hep-G2 cell very low protein expression level (mouse polyclonal anti-human NAPRT antibody) and enzymatic activity (TLC) Homo sapiens
-
6.3.4.21 HL-60 cell
-
 Homo sapiens
-
6.3.4.21 kidney
-
 Mus musculus
-
6.3.4.21 kidney 1.9 kb mRNA detected by Northern blot analysis Mus musculus
-
6.3.4.21 kidney detected by Western blot using mouse polyclonal anti-human NAPRT antibody, robust endogenous activity Rattus norvegicus
-
6.3.4.21 liver
-
 Mus musculus
-
6.3.4.21 liver 1.9 kb mRNA detected by Northern blot analysis Mus musculus
-
6.3.4.21 liver detected by Western blot using mouse polyclonal anti-human NAPRT antibody, robust endogenous activity Rattus norvegicus
-
6.3.4.21 additional information protein not detectable in small intestine Rattus norvegicus
-
6.3.4.21 small intestine
-
 Mus musculus
-
6.3.4.21 small intestine 1.9 kb mRNA detected by Northern blot analysis Mus musculus
-
6.3.4.21 spleen low endogenous activity Rattus norvegicus
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
6.3.4.21 0.0169
-
 +/-0.006 micromol/min/mg, with respect to nicotinic acid, in presence of 1 mM NAD+, 80 ng enzyme, 10-70 microM nicotinamide, 0.3 mM PRPP, 30 min, 37°C, TLC-based quantification Homo sapiens
6.3.4.21 0.0186
-
 +/-0.006 micromol/min/mg, with respect to nicotinic acid, 80 ng enzyme, 10-70 microM nicotinamide, 0.3 mM PRPP, 30 min, 37°C, TLC-based quantification Homo sapiens
6.3.4.21 0.0226
-
 +/-0.0016 micromol/min/mg, with respect to 5-phospho-alpha-D-ribose 1-diphosphate, in presence of 1 mM NAD+, 20 ng enzyme, 40 microM nicotinamide, 0.14-1 microM PRPP, 15 min, 37°C, TLC-based quantification Homo sapiens
6.3.4.21 0.0236
-
 +/-0.0015 micromol/min/mg, with respect to 5-phospho-alpha-D-ribose 1-diphosphate, 20 ng enzyme, 40 microM nicotinamide, 0.14-1 microM PRPP, 15 min, 37°C, TLC-based quantification Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.3.4.21 additional information nicotinamide is not substrate Homo sapiens ?
-
 ?
6.3.4.21 additional information quinolinic acid is not substrate Homo sapiens ?
-
 ?
6.3.4.21 nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + ATP + H2O
-
 Mus musculus beta-nicotinate D-ribonucleotide + diphosphate + ADP + phosphate
-
 ?
6.3.4.21 nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + ATP + H2O
-
 Homo sapiens beta-nicotinate D-ribonucleotide + diphosphate + ADP + phosphate
-
 r
6.3.4.21 nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + ATP + H2O
-
 Rattus norvegicus beta-nicotinate D-ribonucleotide + diphosphate + ADP + phosphate
-
 r
6.3.4.21 nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + ATP + H2O
-
 Mus musculus beta-nicotinate D-ribonucleotide + diphosphate + ADP + phosphate
-
 r
6.3.4.21 nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + ATP + H2O NAPRT is essential for nicotinic acid to increase cellular NAD levels and thus to prevent oxidative stress of the cells Homo sapiens beta-nicotinate D-ribonucleotide + diphosphate + ADP + phosphate
-
 ?
6.3.4.21 nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + ATP + H2O + whole cell lysates, pH 7.5, 37°C, 10 mM MgCl2, 2.5 mM dithiothreitol, 1 mM ATP, 25 microgram bovine serum albumin, with or without NAD+ Homo sapiens beta-nicotinate D-ribonucleotide + diphosphate + ADP + phosphate termination by boiling for 60 seconds, analysis by thin layer chromatography (radiolabeled nicotinic acid) or ESI-MS, no NaMN detectable in Hep-G2 cells r
6.3.4.21 nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + ATP + H2O
-
 Mus musculus BALB/c beta-nicotinate D-ribonucleotide + diphosphate + ADP + phosphate
-
 r

Subunits

EC Number Subunits Comment Organism
6.3.4.21 ? x * 54000 Homo sapiens
6.3.4.21 ? x * 59800, calculated from sequence Homo sapiens
6.3.4.21 homodimer 2 * 55000, non-denaturing polyacrylamide gel electrophoresis of purified recombinant NAPRT in active state Homo sapiens

Synonyms

EC Number Synonyms Comment Organism
6.3.4.21 NA phosphoribosyltransferase
-
 Mus musculus
6.3.4.21 NA phosphoribosyltransferase
-
 Homo sapiens
6.3.4.21 NA phosphoribosyltransferase
-
 Rattus norvegicus
6.3.4.21 NAPRT
-
 Mus musculus
6.3.4.21 NAPRT
-
 Homo sapiens
6.3.4.21 NAPRT
-
 Rattus norvegicus
6.3.4.21 nicotinic acid phosphoribosyltransferase
-
 Mus musculus
6.3.4.21 nicotinic acid phosphoribosyltransferase
-
 Homo sapiens
6.3.4.21 nicotinic acid phosphoribosyltransferase
-
 Rattus norvegicus