Information on EC 6.3.4.21 - nicotinate phosphoribosyltransferase

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].

9030-26-6

-

nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + ATP + H2O

beta-nicotinate D-ribonucleotide + diphosphate + ADP + phosphate

36 entries

nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + GTP + H2O

beta-nicotinate D-ribonucleotide + diphosphate + GDP + phosphate

2 entries

nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + ITP + H2O

beta-nicotinate D-ribonucleotide + diphosphate + IDP + phosphate

2 entries

additional information

?

-

18 entries

nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + ATP + H2O

beta-nicotinate D-ribonucleotide + diphosphate + ADP + phosphate

17 entries

additional information

?

-

ATP

CTP

GTP

ITP

TTP

UTP

Co2+

-

requirement, slight synergism with ATP, more effective than Mg2+ with or without ATP

637754

Fe2+

Sus scrofa

-

slight stimulation, synergism with ATP

Mg2+

MgATP2-

Mn2+

-

requirement, 7.5 mM, no synergism with ATP, more effective than Mg2+ with or without ATP

637754

NaF

Escherichia coli

-

activation

637748

phosphate

5 entries

Zn2+

Sus scrofa

-

slight stimulation

637754

additional information

2 entries

2-hydroxynicotinic acid

-

3-acetylpyridine

-

3-Pyridylcarbinol

-

weak inhibitor

637757

3-Pyridylcarboxaldehyde

Bacillus subtilis

-

637757

5-Fluoronicotinic acid

Bacillus subtilis

-

637757

5-phospho-alpha-D-ribose 1-diphosphate

2 entries

acetyl-CoA

Homo sapiens

Q6XQN6

at 1 mM 40% inhibition of activity (concentrations of substrates 1 mM)

ATP

CoA

at 1 mM 70% inhibition of activity (concentrations of substrates 1 mM)

721763

D-fructose 1,6-bisphosphate

Homo sapiens

Q6XQN6

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)

721763

glyceraldehyde 3-phosphate

Homo sapiens

Q6XQN6

at 1 mM 27% inhibition of activity (concentrations of substrates 1 mM)

GMP

-

IDP

-

IMP

-

ITP

-

KCN

-

Methyl 3-pyridylcarbinol

Bacillus subtilis

-

weak inhibitor

637757

Mg2+

2 entries

MgADP-

Saccharomyces cerevisiae

-

non-competitive to Mg-ATP or Mg-diphosphate

637756

Monoiodoacetic acid

Sus scrofa

-

1 mM and above

637754

N-ethylmaleimide

Sus scrofa

-

0.1 mM and above

637754

NAD+

2 entries

NADH

Sus scrofa

-

weak inhibitor

637754

NADP+

2 entries

NADPH

Sus scrofa

-

weak inhibitor

637754

Nicotinate analogues

2 entries

-

nicotinate mononucleotide

5 entries

nucleoside diphosphates

Bacillus subtilis

-

e.g. ADP or GDP

637757

PCMB

Sus scrofa

-

0.01 mM and above

637754

phosphate

Saccharomyces cerevisiae

-

0.1 M and above

637750

phosphoenolpyruvate

Homo sapiens

Q6XQN6

at 1 mM 33% inhibition of activity (concentrations of substrates 1 mM)

721763

Pyrazine 2-carboxylic acid

Sus scrofa

-

637754

Pyridazine-4,5-dicarboxylic acid

Sus scrofa

-

weak, 10% less activity

637754

succinyl-CoA

Homo sapiens

Q6XQN6

at 1 mM 53% inhibition of activity (concentrations of substrates 1 mM)

tripolyphosphate

-

Urea

-

competitive to nicotinate mononucleotide

637757

UTP

Bos taurus

-

weak

637749

additional information

8 entries

-

ATP

2 entries

GSH

Escherichia coli

-

activation

637748

tripolyphosphate

Bacillus subtilis

-

activation, can replace ATP

637757

additional information

7 entries

-

0.00022 - 48

5-phospho-alpha-D-ribose 1-diphosphate

17 entries

0.07 - 0.4

ATP

5 entries

0.34

GTP

Bacillus subtilis

-

637757

1.16

ITP

Salmonella enterica subsp. enterica serovar Typhimurium

-

good alternative substrate for ATP

637758

0.0005 - 0.4

nicotinate

15 entries

0.0127 - 0.0155

nicotinic acid

2 entries

0.01 - 0.36

5-phospho-alpha-D-ribose 1-diphosphate

2 entries

0.012 - 0.25

nicotinate

2 entries

additional information

2 entries

-

0.452 - 9.42

5-phospho-alpha-D-ribose 1-diphosphate

2 entries

0.271 - 9.16

nicotinate

2 entries

0.85

CoA

Homo sapiens

pH 7.5, 37°C

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, 37°C, TLC-based quantification

680797

0.0186

Homo sapiens

Q6XQN6

+/-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

680797

0.0226

Homo sapiens

Q6XQN6

+/-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

680797

0.0236

Homo sapiens

Q6XQN6

+/-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

0.052

-

0.08

-

recombinant enzyme from Escherichia coli strain BA203, pH and temperature not specified in the publication

727386

0.095

Sus scrofa

-

crude extract

637754

0.7

2 entries

0.74

Escherichia coli

-

recombinant enzyme from Escherichia coli strain BA204, pH and temperature not specified in the publication

1.01

10.54

12.24

-

recombinant enzyme from Escherichia coli strain BA207, pH and temperature not specified in the publication

726892

160.5

Sus scrofa

-

DEAE-Sephadex fraction

637754

17.31

Escherichia coli

-

recombinant enzyme from Escherichia coli strain BA206, pH and temperature not specified in the publication

727386

18.74

Escherichia coli

P18133

recombinant enzyme from Escherichia coli strain BA016, pH and temperature not specified in the publication

726780

2.3

Saccharomyces cerevisiae

-

637750

23.72

Escherichia coli

-

recombinant enzyme from Escherichia coli strain BA207, pH and temperature not specified in the publication

727386

3

Salmonella enterica subsp. enterica serovar Typhimurium

-

637761

421.2

Sus scrofa

-

Sephacryl fraction

637754

457.3

Sus scrofa

-

phenyl-Sepharose fraction

637754

6.5 - 8

Homo sapiens

-

broad, 0.2 M potassium phosphate buffer or 0.05 M Tris-Cl

637752

7.2

Bos taurus

-

broad, phosphorolysis

637749

7.3 - 7.4

Sus scrofa

-

637754

7.5 - 8.5

Saccharomyces cerevisiae

-

broad

637750

8.5

2 entries

additional information

2 entries

5.5 - 10

Saccharomyces cerevisiae

-

about half-maximal activity at pH 5.5 and 10

637750

5.5 - 8

Bos taurus

-

about half-maximal activity at pH 5.5 and about 85% of maximal activity at pH 8

637749

6.5 - 9

Bacillus subtilis

-

637757

additional information

37

SB19

-

Bacillus subtilis SB19

SB19

-

beef

-

-

-

BALB/c

UniProt

Mycobacterium tuberculosis

3 entries

Mycobacterium tuberculosis H37Rv

3 entries

Oryza sativa Japonica Group

-

UniProt

-

-

Rattus norvegicus male Wistar

male Wistar strain

637754

-

Saccharomyces cerevisiae

7 entries

Salmonella enterica subsp. enterica serovar Typhimurium

2 entries

Sus scrofa

2 entries

Thermoplasma acidophilum

-

Uniprot

-

UniProt

Trichoderma reesei ATCC 56765

-

UniProt

-

weak expression

-

-

weak expression

745944

Caki-2 cell

Homo sapiens

-

weak expression

-

cell suspension culture

-

-

Phoenix cell

-

-

very low protein expression level (mouse polyclonal anti-human NAPRT antibody) and enzymatic activity (TLC)

-

-

-

-

-

-

additional information

Highest Expressing Human Cell Lines

Filter by:

Cell Line Links

Gene Links

physiological function

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

680797

120000

Sus scrofa

-

liver, SDS-PAGE

637754

43000

2 entries

45000

3 entries

45500

Salmonella enterica subsp. enterica serovar Typhimurium

-

mutant enzyme, SDS-PAGE

637759

45529

Salmonella enterica subsp. enterica serovar Typhimurium

-

1 * 45529, gel filtration

637759

45530

Salmonella enterica subsp. enterica serovar Typhimurium

P22253

predicted from DNA sequence

637763

45540

Salmonella enterica subsp. enterica serovar Typhimurium

-

SDS-PAGE

637759

45550

Salmonella enterica subsp. enterica serovar Typhimurium

-

mutant enzyme, SDS-PAGE

46000

x * 46000, SDS-PAGE

51000

52000

overexpressed, His-tagged NAPRT in Hep-G2 cells detected in Western blot using mouse polyclonal anti-human NAPRT antibody

680797

54000

Homo sapiens

Q6XQN6

purified recombinant NAPRT

680797

55000

Homo sapiens

Q6XQN6

2 * 55000, non-denaturing polyacrylamide gel electrophoresis of purified recombinant NAPRT in active state

680797

59800

Homo sapiens

Q6XQN6

calculated from the deduced protein sequence

680797

63000

Sus scrofa

-

2 * 63000, SDS-PAGE

637754

86000

Homo sapiens

-

gel filtration

637752

87000

Homo sapiens

Q6XQN6

gel filtration, predicted value of monomer: 59249 Da

721763

?

5 entries

dimer

Sus scrofa

-

2 * 63000, SDS-PAGE

637754

hexamer

Thermoplasma acidophilum

Q9HJ28

6 * 43000, (alphabeta)3, SDS-PAGE and crystal structure analysis

659469

homodimer

Homo sapiens

Q6XQN6

2 * 55000, non-denaturing polyacrylamide gel electrophoresis of purified recombinant NAPRT in active state

680797

monomer

2 entries

additional information

2 entries

by molecular replacement at a resolution of 2.9 A, in its ligand-free form. The enzyme consists of two domains and functions as a dimer with the active site located at the interface of the monomers. NaPRTase acks a tunnel that, in nicotinamide phosphoribosyltransferase EC 2.4.2.12, represents the binding site of inhibitor FK866

Homo sapiens

Q6XQN6

736115

recombinant selenomethionine-labeled enzyme, 10-30 mg/ml, in 10 mM Tris, pH 7.8, 150 mM NaCl, nanodroplet vapour diffusion method, 4°C, 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

Saccharomyces cerevisiae

P39683

660524

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

Thermoplasma acidophilum

D19A

activity completely abolished

721763

D288A

Homo sapiens

Q6XQN6

about 10% of wild type activity

721763

G169A

Homo sapiens

Q6XQN6

about 40% of wild type activity

721763

G209A

Homo sapiens

Q6XQN6

about 20% of wild type activity

721763

G379A

Homo sapiens

Q6XQN6

activity completely abolished

H213A

N357A

about 80% of wild type activity

721763

R318A

Homo sapiens

Q6XQN6

activity strictly dependent on the presence of ATP

721763

S381A

Homo sapiens

Q6XQN6

about 40% of wild type activity

721763

T380A

Homo sapiens

Q6XQN6

about 50% of wild type activity

721763

Y21A

Homo sapiens

Q6XQN6

activity strictly dependent on the presence of ATP

721763

A323V

Mycobacterium tuberculosis

P9WJI9, P9WJI7

mutation identified from whole-genome sequenced data

775420

F286V

Mycobacterium tuberculosis

P9WJI9, P9WJI7

mutation identified from whole-genome sequenced data

775420

G127S

2 entries

G429A

Mycobacterium tuberculosis

P9WJI9, P9WJI7

mutation has stabilizing effects on the protein structure

775420

P447S

2 entries

S120L

Mycobacterium tuberculosis

P9WJI9, P9WJI7

mutation localized in the loop region, leads to a gain in flexibility in the surrounding residues

775420

A323V

Mycobacterium tuberculosis H37Rv

-

mutation identified from whole-genome sequenced data

-

F286V

Mycobacterium tuberculosis H37Rv

-

mutation identified from whole-genome sequenced data

-

G127S

Mycobacterium tuberculosis H37Rv

-

mutation identified from whole-genome sequenced data

-

P447S

2 entries

S120L

Mycobacterium tuberculosis H37Rv

-

mutation localized in the loop region, leads to a gain in flexibility in the surrounding residues

-

P21L

Oryza sativa Japonica Group

Q851M0

mutation identified in a strain with early leaf sensescence, mutation causes the expression of the gene at the transcriptional level to decline by 40% in fully expanded leaves. Leaf nicotinate and nicotinamide contents are elevated, while NAD levels are reduced. Leaf tissue of P21L exhibits significant DNA fragmentation and H2O2 accumulation, along with up-regulation of genes associated with senescence. The mutant also shows reduced expression of SIR2-like genes SRT1 and SRT2 and increased acetylation of histone H3K9

746114

additional information

6 entries

5 - 10

Homo sapiens

-

stable, 0.2 M potassium phosphate buffer

637752

6.5

Bos taurus

-

inactivation below, phosphorolysis

637749

8.3

Salmonella enterica subsp. enterica serovar Typhimurium

P22253

native enzyme, unstable

637763

50

Saccharomyces cerevisiae

-

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

637750

additional information

3 entries

50% activity recovered at salt concentrations of 50 mM, inactivation with salt concentrations below 20 mM, 20-35% glycerol partially protects

Homo sapiens

-

637752

dialysis against 50 mM Tris-HCl buffer at pH 7.3 in the absence of potassium phosphate, significant loss of activity

2 entries

first order activity loss in the presence of 1/200 trypsin/enzyme

Salmonella enterica subsp. enterica serovar Typhimurium

-

637759, 637760

glycerol, 20-35%, stabilizes during dialysis against low buffer concentrations, more highly purified preparations require higher glycerol concentrations

Homo sapiens

-

637752

heat, unstable to, substrates protect

Saccharomyces cerevisiae

-

637750

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

Salmonella enterica subsp. enterica serovar Typhimurium

-

637760

sensitive to trypsin, partial proteolysis, 4fold increase in t1/2 for inactivation in the presence of 1 mM MgATP2-

Salmonella enterica subsp. enterica serovar Typhimurium

-

637759

-12°C, partially purified enzyme preparation, t1/2: at least 3 months

Bos taurus

-

637749

-25°C, at least 1 month

Saccharomyces cerevisiae

-

637750

-70°C, t1/2: 3 months

Homo sapiens

-

637752

-70ºC, no significantly decreased activity for up to 3 years, no significantly decreased after 8 years

Homo sapiens

-

637762

4ºC, as 70% saturated ammonium sulfate suspensions in 200 mM phosphate buffer, pH 8, 10% glycerol and 5 mM DTT

Salmonella enterica subsp. enterica serovar Typhimurium

-

637759, 637760, 637761

t1/2: at least 2 months

Salmonella enterica subsp. enterica serovar Typhimurium

-

637761

-

2 entries

ammonium hydroxide precipitation, chromatography on DEAE-Sephadex, Sephacryl and phenyl-Sepharose

Sus scrofa

-

637754

ammonium sulfate fractionation, chromatography on DEAE-cellulose, chromatography on Sephadex, affinity column chromatography and hydroxylapatite column chromatography, 30000fold purified

Homo sapiens

-

637752

bacteria disruption by alumina grinding, protamine sulfate precipitation and chromatography on DEAE-cellulose

Escherichia coli

-

637748

bacterial suspension disruption and centrifugation, chromatography on Q-Sepharose and phenyl-Sepharose

Salmonella enterica subsp. enterica serovar Typhimurium

-

637760

bacterial suspension disruption and centrifugation, chromatography on Sephadex-G, DEAE-Toyopearl, Cibacron Blue and Sephacryl gel filtration

Salmonella enterica subsp. enterica serovar Typhimurium

-

637761

extraction, ammonium sulfate fractionation and chromatography on DEAE-cellulose

Bos taurus

-

637749

extraction, ammonium sulfate fractionation, chromatography on DEAE-cellulose and hydroxylapatite, 1000fold purified

Saccharomyces cerevisiae

-

637750

extraction, fractionation chromatography on phosphocellulose, hydroxyapatite, blue Sephadex and DEAE-cellulose

Saccharomyces cerevisiae

-

637753

heparinised blood lysates

-

homogenates

-

immobilized metal ion affinity chromatography (Ni2+)

Homo sapiens

Q6XQN6

721763

nickel chelate His-bind resin after bacterial expression

Homo sapiens

Q6XQN6

680797

partial, extraction, ammonium sulfate fractionation and chromatography on DEAE-cellulose, 60fold purified

Bacillus subtilis

-

637757

recombinant N-terminally tagged selenomethionine-labeled enzyme variant from Escherichia coli strain DL41

Saccharomyces cerevisiae

P39683

660524

recombinant tagged wild-type and selenomethionine-labeled fusion enzymes from Escherichia coli to homogeneity by high-trap chelating nickel affinity chromatography

Thermoplasma acidophilum

Q9HJ28

659469

-

Oryza sativa Japonica Group

Q851M0

746114

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

Q8CC86

680797

expressed in Escherichia coli NZN111

Escherichia coli

P18133

722141

expressed in Escherichia coli strains BA002, BA014, BA015, and BA016

Escherichia coli

P18133

726780

expressed in Escherichia coli strains BA002, BA014, BA026, and BA207

Escherichia coli

-

726892

expressed in Escherichia coli strains BA203, BA204, BA206, and BA209

Escherichia coli

-

727386

expression in Escherichia coli

Saccharomyces cerevisiae

P39683

637759

expression in Escherichia coli as His6-tagged protein

Homo sapiens

Q6XQN6

680797

expression of N-terminally tagged enzyme as selenomethionine-labeled variant in Escherichia coli strain DL41

Saccharomyces cerevisiae

P39683

660524

expression of pncB gene on a multicopy pUC19 plasmid in Salmonella typhimurium

Salmonella enterica subsp. enterica serovar Typhimurium

-

637761

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

Thermoplasma acidophilum

Q9HJ28

659469

His-tagged protein expressed in Escherichia coli BL21

Homo sapiens

Q6XQN6

721763

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

Q6XQN6

680797

presence of more than one NAPRT transcript in most tissues, all with identical 3' end sequence. Some NAPRT transcripts appear to be tissuespecific. Brain shows the highest number of alternative transcripts. Several modulators of the NAPRT gene expression are involved, i.e. mutations in transcription factor binding sites, promoter methylation and alternative splicing

-

degradation

medicine

ExplorEnz (official portal for IUBMB Enzyme Nomenclature)

Expasy Enzyme Nomenclature Database

KEGG

SABIO-RK

NCBI: PubMed, Protein, Nucleotide, Structure, Gene, OMIM

Enzyme Nomenclature (alternative site)

UniProt

PDB

PROSITE Database of protein families and domains

InterPro (database of protein families, domains and functional sites)