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 |
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 |
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 |
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 | - |
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 |
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 |
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 |
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 | - |
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 |
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 | - |
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 |
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 |
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 |
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 |