Information on EC 6.3.4.21 - nicotinate phosphoribosyltransferase

Word Map on EC 6.3.4.21
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea

EC NUMBER
COMMENTARY hide
6.3.4.21
-
RECOMMENDED NAME
GeneOntology No.
nicotinate phosphoribosyltransferase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + ATP + H2O = beta-nicotinate D-ribonucleotide + diphosphate + ADP + phosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pentosyl group transfer
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Metabolic pathways
-
-
NAD salvage pathway I
-
-
Nicotinate and nicotinamide metabolism
-
-
pyridine nucleotide cycling (plants)
-
-
NAD metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
5-phospho-alpha-D-ribose 1-diphosphate:nicotinate ligase (ADP, diphosphate-forming)
The enzyme, which is involved in pyridine nucleotide recycling, can form beta-nicotinate D-ribonucleotide and diphosphate from nicotinate and 5-phospho-alpha-D-ribose 1-diphosphate (PRPP) in the absence of ATP. However, when ATP is available the enzyme is phosphorylated resulting in a much lower Km for nicotinate. The phospho-enzyme is hydrolysed during the transferase reaction, regenerating the low affinity form. The presence of ATP shifts the products/substrates equilibrium from 0.67 to 1100 [4].
CAS REGISTRY NUMBER
COMMENTARY hide
9030-26-6
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SB19
-
-
Manually annotated by BRENDA team
SB19
-
-
Manually annotated by BRENDA team
beef
-
-
Manually annotated by BRENDA team
BALB/c
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
male Wistar strain
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + ATP + H2O
beta-nicotinate D-ribonucleotide + diphosphate + ADP + phosphate
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + ATP + H2O
beta-nicotinate D-ribonucleotide + diphosphate + ADP + phosphate
show the reaction diagram
additional information
?
-
P39683
molecular evolution mechanism
-
-
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
-
requirement, slight synergism with ATP, more effective than Mg2+ with or without ATP
Fe2+
-
slight stimulation, synergism with ATP
MgATP2-
Mn2+
-
requirement, 7.5 mM, no synergism with ATP, more effective than Mg2+ with or without ATP
NaF
-
activation
phosphate
Zn2+
-
slight stimulation
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3-acetylpyridine
-
-
3-Pyridylcarbinol
-
weak inhibitor
3-Pyridylcarboxaldehyde
-
-
5-Fluoronicotinic acid
-
-
5-phospho-alpha-D-ribose 1-diphosphate
acetyl-CoA
-
at 1 mM 40% inhibition of activity (concentrations of substrates 1 mM)
CoA
-
at 1 mM 70% inhibition of activity (concentrations of substrates 1 mM)
D-fructose 1,6-bisphosphate
-
at 1 mM 36% inhibition of activity (concentrations of substrates 1 mM)
diphosphate
-
-
GDP
-
-
glutaryl-CoA
-
at 1 mM 42% inhibition of activity (concentrations of substrates 1 mM)
glyceraldehyde 3-phosphate
-
at 1 mM 27% inhibition of activity (concentrations of substrates 1 mM)
GMP
-
-
IDP
-
-
IMP
-
-
ITP
-
-
Methyl 3-pyridylcarbinol
-
weak inhibitor
MgADP-
-
non-competitive to Mg-ATP or Mg-diphosphate
Monoiodoacetic acid
-
1 mM and above
N-ethylmaleimide
-
0.1 mM and above
NADH
-
weak inhibitor
NADP+
NADPH
-
weak inhibitor
Nicotinate analogues
-
nicotinate mononucleotide
nucleoside diphosphates
-
e.g. ADP or GDP
PCMB
-
0.01 mM and above
phosphate
-
0.1 M and above
phosphoenolpyruvate
-
at 1 mM 33% inhibition of activity (concentrations of substrates 1 mM)
Pyrazine 2-carboxylic acid
-
-
Pyridazine-4,5-dicarboxylic acid
-
weak, 10% less activity
succinyl-CoA
-
at 1 mM 53% inhibition of activity (concentrations of substrates 1 mM)
tripolyphosphate
-
-
Urea
-
competitive to nicotinate mononucleotide
UTP
-
weak
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
GSH
-
activation
tripolyphosphate
-
activation, can replace ATP
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00022 - 48
5-phospho-alpha-D-ribose 1-diphosphate
0.07 - 0.4
ATP
0.34
GTP
-
-
1.16
ITP
-
good alternative substrate for ATP
0.02
Mg-5-phospho-alpha-D-ribose 1-diphosphate
-
in the coupled reaction, 200fold lower than in the uncoupled reaction
0.0005 - 0.4
nicotinate
0.0127 - 0.0155
nicotinic acid
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.01 - 0.36
5-phospho-alpha-D-ribose 1-diphosphate
0.012 - 0.25
nicotinate
additional information
additional information
Saccharomyces cerevisiae
P39683
-
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.452 - 9.42
5-phospho-alpha-D-ribose 1-diphosphate
158
0.271 - 9.16
nicotinate
1029
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.85
CoA
Homo sapiens
-
pH 7.5, 37C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0012
-
-
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, 37C, TLC-based quantification
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, 37C, TLC-based quantification
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, 37C, TLC-based quantification
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, 37C, TLC-based quantification
0.052
-
-
0.08
-
recombinant enzyme from Escherichia coli strain BA203, pH and temperature not specified in the publication
0.095
-
crude extract
0.7
-
recombinant enzyme from Escherichia coli strain BA002, pH and temperature not specified in the publication
0.74
-
recombinant enzyme from Escherichia coli strain BA204, pH and temperature not specified in the publication
10.54
-
recombinant enzyme from Escherichia coli strain BA014, pH and temperature not specified in the publication
12.24
-
recombinant enzyme from Escherichia coli strain BA207, pH and temperature not specified in the publication
17.31
-
recombinant enzyme from Escherichia coli strain BA206, pH and temperature not specified in the publication
18.74
-
recombinant enzyme from Escherichia coli strain BA016, pH and temperature not specified in the publication
23.72
-
recombinant enzyme from Escherichia coli strain BA207, pH and temperature not specified in the publication
160.5
-
DEAE-Sephadex fraction
421.2
-
Sephacryl fraction
457.3
-
phenyl-Sepharose fraction
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 8
-
broad, 0.2 M potassium phosphate buffer or 0.05 M Tris-Cl
7.2
-
broad, phosphorolysis
7.3 - 7.4
-
-
7.5 - 8.5
-
broad
additional information
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 8
-
about half-maximal activity at pH 5.5 and about 85% of maximal activity at pH 8
5.5 - 10
-
about half-maximal activity at pH 5.5 and 10
additional information
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
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
Manually annotated by BRENDA team
-
very low protein expression level (mouse polyclonal anti-human NAPRT antibody) and enzymatic activity (TLC)
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Agrobacterium fabrum (strain C58 / ATCC 33970)
Enterococcus faecalis (strain ATCC 700802 / V583)
Enterococcus faecalis (strain ATCC 700802 / V583)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Thermoplasma acidophilum (strain ATCC 25905 / DSM 1728 / JCM 9062 / NBRC 15155 / AMRC-C165)
Thermoplasma acidophilum (strain ATCC 25905 / DSM 1728 / JCM 9062 / NBRC 15155 / AMRC-C165)
Thermoplasma acidophilum (strain ATCC 25905 / DSM 1728 / JCM 9062 / NBRC 15155 / AMRC-C165)
Thermoplasma acidophilum (strain ATCC 25905 / DSM 1728 / JCM 9062 / NBRC 15155 / AMRC-C165)
Vibrio cholerae serotype O1 (strain ATCC 39315 / El Tor Inaba N16961)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
46000
-
x * 46000, SDS-PAGE
52000
-
overexpressed, His-tagged NAPRT in Hep-G2 cells detected in Western blot using mouse polyclonal anti-human NAPRT antibody
54000
-
purified recombinant NAPRT
55000
-
2 * 55000, non-denaturing polyacrylamide gel electrophoresis of purified recombinant NAPRT in active state
59800
-
calculated from the deduced protein sequence
63000
-
2 * 63000, SDS-PAGE
86000
-
gel filtration
87000
-
gel filtration, predicted value of monomer: 59249 Da
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, 37C
120000
-
liver, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
-
2 * 63000, SDS-PAGE
hexamer
6 * 43000, (alphabeta)3, SDS-PAGE and crystal structure analysis
homodimer
-
2 * 55000, non-denaturing polyacrylamide gel electrophoresis of purified recombinant NAPRT in active state
monomer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant selenomethionine-labeled enzyme, 10-30 mg/ml, in 10 mM Tris, pH 7.8, 150 mM NaCl, nanodroplet vapour diffusion method, 4C, reservoir solution contains 6-16% PEG 5000 monomethyl ether, 0.06 M MES, and 0.04 M NaMES, pH 6.0, usage of 15-20% ethylene glycol as cryoprotectant, X-ray structure determination and analysis at 1.75 A resolution
recombinant detagged wild-type and selenomethionine-labeled enzymes, hanging drop vapour diffusion method, room temperature, 0.001 ml protein solution mixed with equal volume of well solution containing 0.2 M ammonium acetate, 0.1 M Tris-HCl, pH 8.5, 45% 2-methyl-2,4-pentanediol, 1 week, crystallization condition screening using sitting drop vapour diffusion at room temperature, iridium complexed, X-ray diffraction structure determination and analysis at 2.8 A resolution
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 10
-
stable, 0.2 M potassium phosphate buffer
637752
6.5
-
inactivation below, phosphorolysis
637749
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50
-
5 min: 95% loss of activity, MgATP2-, Mg-phosphoribose diphosphate, ATP, GTP or nicotinic acid protect, with 27%, 37%, 52%, 54% or 61% loss of activity, respectively. 10 min: 97% loss of activity, MgATP2-, Mg-5-phospho-alpha-D-ribose 1-diphosphate, ATP, GTP or nicotinic acid protect, with 43%, 52%, 60%, 63% or 69% loss of activity, respectively
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
50% activity recovered at salt concentrations of 50 mM, inactivation with salt concentrations below 20 mM, 20-35% glycerol partially protects
-
dialysis against 50 mM Tris-HCl buffer at pH 7.3 in the absence of potassium phosphate, significant loss of activity
first order activity loss in the presence of 1/200 trypsin/enzyme
-
glycerol, 20-35%, stabilizes during dialysis against low buffer concentrations, more highly purified preparations require higher glycerol concentrations
-
heat, unstable to, substrates protect
-
rapidly inactivates the ATPase and nicotinic acid mononucleotide synthesis activities with cleavages at Arg-384 and Lys-374, ATP and 5-phospho-alpha-D-ribose 1-diphosphate increase the t1/2 for inactivation 8fold and 4fold, respectively, and 32fold, together
-
sensitive to trypsin, partial proteolysis, 4fold increase in t1/2 for inactivation in the presence of 1 mM MgATP2-
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-12C, partially purified enzyme preparation, t1/2: at least 3 months
-
-25C, at least 1 month
-
-70C, t1/2: 3 months
-
-70C, no significantly decreased activity for up to 3 years, no significantly decreased after 8 years
-
4C, as 70% saturated ammonium sulfate suspensions in 200 mM phosphate buffer, pH 8, 10% glycerol and 5 mM DTT
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; nickel chelate His-bind resin after bacterial expression
-
ammonium hydroxide precipitation, chromatography on DEAE-Sephadex, Sephacryl and phenyl-Sepharose
-
ammonium sulfate fractionation, chromatography on DEAE-cellulose, chromatography on Sephadex, affinity column chromatography and hydroxylapatite column chromatography, 30000fold purified
-
bacteria disruption by alumina grinding, protamine sulfate precipitation and chromatography on DEAE-cellulose
-
bacterial suspension disruption and centrifugation, chromatography on Q-Sepharose and phenyl-Sepharose
-
bacterial suspension disruption and centrifugation, chromatography on Sephadex-G, DEAE-Toyopearl, Cibacron Blue and Sephacryl gel filtration
-
extraction, ammonium sulfate fractionation and chromatography on DEAE-cellulose
-
extraction, ammonium sulfate fractionation, chromatography on DEAE-cellulose and hydroxylapatite, 1000fold purified
-
extraction, fractionation chromatography on phosphocellulose, hydroxyapatite, blue Sephadex and DEAE-cellulose
-
heparinised blood lysates
-
homogenates
-
immobilized metal ion affinity chromatography (Ni2+)
-
partial, extraction, ammonium sulfate fractionation and chromatography on DEAE-cellulose, 60fold purified
-
recombinant N-terminally tagged selenomethionine-labeled enzyme variant from Escherichia coli strain DL41
recombinant tagged wild-type and selenomethionine-labeled fusion enzymes from Escherichia coli to homogeneity by high-trap chelating nickel affinity chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
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
expressed in Escherichia coli NZN111
-
expressed in Escherichia coli strains BA002, BA014, BA015, and BA016
-
expressed in Escherichia coli strains BA002, BA014, BA026, and BA207
-
expressed in Escherichia coli strains BA203, BA204, BA206, and BA209
-
expression in Escherichia coli
-
expression in Escherichia coli as His6-tagged protein; 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
-
expression of N-terminally tagged enzyme as selenomethionine-labeled variant in Escherichia coli strain DL41
expression of pncB gene on a multicopy pUC19 plasmid in Salmonella typhimurium
-
expression of the enzyme as His6-tagged-maltose-binding fusion protein containing a tobacco etch virus protease cleavage site in Escherichia coli strain BL21(DE3) for the wild-type, and in strain B834(DE3) for a selenomethionine-labeled variant
His-tagged protein expressed in Escherichia coli BL21
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D19A
-
activity completely abolished
D288A
-
about 10% of wild type activity
G169A
-
about 40% of wild type activity
G209A
-
about 20% of wild type activity
G379A
-
activity completely abolished
H213A
-
about 50% of wild type activity
N357A
-
about 80% of wild type activity
R318A
-
activity strictly dependent on the presence of ATP
S381A
-
about 40% of wild type activity
T380A
-
about 50% of wild type activity
Y21A
-
activity strictly dependent on the presence of ATP
Show AA Sequence (11623 entries)
Longer loading times are possible. Please use the Sequence Search for a specific query.