Information on EC 2.4.2.12 - nicotinamide phosphoribosyltransferase

Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
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

EC NUMBER
COMMENTARY
2.4.2.12
-
RECOMMENDED NAME
GeneOntology No.
nicotinamide phosphoribosyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
nicotinamide D-ribonucleotide + diphosphate = nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
show the reaction diagram
-
-
-
-
nicotinamide D-ribonucleotide + diphosphate = nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
show the reaction diagram
mechanism
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
pentosyl group transfer
-
-
-
-
pentosyl group transfer
Q80Z29
-
pentosyl group transfer
-
-
pentosyl group transfer
Q99KQ4
-
PATHWAY
KEGG Link
MetaCyc Link
NAD biosynthesis III
-
Nicotinate and nicotinamide metabolism
-
SYSTEMATIC NAME
IUBMB Comments
nicotinamide-D-ribonucleotide:diphosphate phospho-alpha-D-ribosyltransferase
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
NAmPRTase
-
-
-
-
Nampt
-
-
Nampt
-
-
Nampt
Q99KQ4
-
Nampt
Q80Z29
-
Nampt/PBEF/Visfatin
P43490
-
Nampt/visfatin
Q80Z29
-
NAPRT
-
-
nicotinamide mononucleotide pyrophosphorylase
-
-
-
-
nicotinamide mononucleotide synthetase
-
-
-
-
nicotinamide phosphoribosyl transferase
-
-
nicotinamide phosphoribosyltransferase
-
-
nicotinamide phosphoribosyltransferase
P43490
-
nicotinamide phosphoribosyltransferase
-
-
nicotinamide phosphoribosyltransferase
Q99KQ4
-
nicotinamide phosphoribosyltransferase
-
-
nicotinamide phosphoribosyltransferase
Q80Z29
-
nicotinamide phosphoribosyltransferase/Visfatin
-
does not catalyze nicotinamide mononucleotide formation in blood plasma
NMN pyrophosphorylase
-
-
-
-
NMN synthetase
-
-
-
-
NMPRTase
-
-
NMPRTase
P43490
-
NMPRTase
-
-
PBEF
-
-
-
-
PBEF
-
-
PBEF
P43490
-
PBEF
-
-
PBEF
Q99KQ4
-
phosphoribosyltransferase, nicotinamide
-
-
-
-
pre-B cell colony enhancing factor
Q80Z29
-
pre-B cell colony enhancing factor(PBEF)
-
-
pre-B cell colony-enhancing factor (PBEF)
-
-
pre-B cell colony-enhancing factor 1
Q80Z29
-
pre-B-cell colony enhancing factor
P43490
-
pre-B-cell colony-enhancing factor
-
-
-
-
pre-B-cell colony-enhancing factor
P43490
-
pre-B-cell colony-enhancing factor
Q99KQ4
-
pre-B-cell colony-enhancing factor
-
-
Visfatin
-
-
Visfatin
P43490
-
Visfatin
-
-
Visfatin
Q99KQ4
-
Visfatin
Q80Z29
-
CAS REGISTRY NUMBER
COMMENTARY
9030-27-7
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
-
Q99KQ4
SwissProt
Manually annotated by BRENDA team
identified as pre-B-cell colony-enhancing factor
-
-
Manually annotated by BRENDA team
male DBA/1 mice, naive or with collagen-induced arthritis (CIA) after immunization with type II collagen, and male and female C57BL/6 mice
Q99KQ4
SwissProt
Manually annotated by BRENDA team
no activity in Picea glauca
suspension cultured cells
-
-
Manually annotated by BRENDA team
male Sprague-Dawley
-
-
Manually annotated by BRENDA team
male Wistar rats, six-months-old, develop diet-induced hyperphagia and obesity, male obese (fa/fa) Zucker rats, three-months-old, display genetic obesity due to deficit in leptin receptors, and male lean Zucker rats, three-month-old
SWissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
malfunction
-
implicated in the pathogenesis of a number of human diseases or conditions such as acute lung injury, aging, atherosclerosis, cancer, diabetes, rheumatoid arthritis and sepsis
physiological function
-
cardiac-specific overexpression of Nampt in transgenic mice increases NAD+ content in the heart, prevents downregulation of Nampt, and reduces the size of myocardial infarction and apoptosis in response to prolonged ischemia and ischemia/reperfusion. Upregulation of Nampt significantly increases NAD+ and ATP concentrations, whereas downregulation of Nampt significantly decreases them. Expression of Nampt in the heart is significantly decreased by ischemia, ischemia/reperfusion and pressure overload. Downregulation of Nampt increases caspase 3 cleavage, cytochrome c release, and TUNEL-positive cells, which are inhibited in the presence of Bcl-xL, but do not increase hairpin 2-positive cells, suggesting that endogenous Nampt negatively regulates apoptosis but not necrosis. Downregulation of Nampt also impairs autophagic flux
physiological function
-
growth factor, cytokine and nicotinamide phosphoribosyltransferase
physiological function
-
required for de novo lipogenesis in tumor cells
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ADP + ATP
ATP + ADP
show the reaction diagram
-
isotope exchange reaction (in presence of radiolabeled ADP and ATP but in absence of other reactants, meaning without NMN synthesis) requires high-energy phosphorylated NAMPT
-
-
r
ATP + H2O
ADP + phosphate
show the reaction diagram
-
DELTAG = -7.3, in presence of NAMPT slow steady-state reaction after initial burst, ATPase activity in absence of other substrates, but can be modulated by addition of single substrates and mixtures
-
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
-
-
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
-
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
-
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
-
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
-
-
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
-
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
-
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
-
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
-
-
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
Q99KQ4
-
-
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
specific for nicotinamide, no substrates: thymine, 5-bromouracil, nicotinic acid adenine dinucleotide
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
beta-nicotinamide
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
first step in NAD synthesis from nicotinamide
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
pre-B-cell colony enhancing factor
-
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
key enzyme in the regulation of NAD+ biosynthesis from the natural precursor nicotinamide
-
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
Q99KQ4
key enzyme in the regulation of NAD+ biosynthesis, regulates the silent regulator protein 2, i.e. Sir2, activity, the enzyme has a metabolic regulatory function by up- and downregulation of proteins, overview
-
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
reaction is reversible as dictated by the equilibrium constant K, [NMN][PPi]/([NM][PRPP]) of 0.14, which agrees well with the ratio of second-order rate constants for forward and backward reactions, K of0.16
-
-
r
nicotinamide + 5-phosphoribosyl 1-diphosphate
nicotinamide mononucleotide + diphosphate
show the reaction diagram
-
-
-
-
?
nicotinamide + 5-phosphoribosyl 1-diphosphate
nicotinamide mononucleotide + diphosphate
show the reaction diagram
-
-
-
-
?
nicotinamide + alpha-D-5-phosphoribosyl 1-diphosphate
nicotinamide mononucleotide + diphosphate
show the reaction diagram
-
-
-
-
r
nicotinamide + alpha-D-5-phosphoribosyl 1-diphosphate
nicotinamide mononucleotide + diphosphate
show the reaction diagram
Q99KQ4
nicotinamide clearance
NMN synthesis is rate-limiting step for NAD+ synthesis
-
r
nicotinamide + alpha-D-5-phosphoribosyl 1-diphosphate
nicotinamide mononucleotide + diphosphate
show the reaction diagram
-
nicotinamide clearance
NMN synthesis is rate-limiting step for NAD+ synthesis
-
r
nicotinamide + alpha-D-5-phosphoribosyl 1-diphosphate
nicotinamide mononucleotide + diphosphate
show the reaction diagram
-
DELTAG = 1.1 kcal/mol, Keq = 0.15, unfavourable product-to-substrate ratio
equilibrium was established after 3 h in presence of 100 microM PRPP, 100 microM diphosphate, 5 microM nicotinamide mononucleotide, 0.6 microM NAMPT
-
r
nicotinamide + alpha-D-5-phosphoribosyl 1-diphosphate + ATP + H2O
nicotinamide mononucleotide + diphosphate + ADP + phosphate
show the reaction diagram
-
DELTAG = -1.0 kcal/mol, Keq = 5.0, reversion of the preestablished equilibrium in a non-ATP-coupled reaction, thermodynamic switch of the product-to-substrate ratio towards NMN production compared to the non-ATP-coupled reaction with an energy difference of -2.1 kcal/mol (meaning only 2.1 kcal/mol of 7.3 kcal/mol of the energy from ATP hydrolysis coupled to NMN formation), formation or hydrolysis of high-energy phosphorylated NAMPT may be rate limiting for overall ATP-coupled NMN synthesis
in presence of 2-2.5 mM ATP: 1 mol ADP per NMN formed (stoichiometric coupling ratio, R = 1), under optimal conditions (2 microM nicotinamide, 100 microM PRPP, 2.5 mM ATP, 5 mM Mg2+): R increases linearly with increasing diphosphate concentration
-
r
nicotinamide + alpha-D-5-phosphoribosyl-1-diphosphate
nicotinamide mononucleotide + diphosphate
show the reaction diagram
-
-
-
-
r
nicotinamide + alpha-D-5-phosphoribosyl-1-diphosphate
nicotinamide mononucleotide + inorganic diphosphate
show the reaction diagram
Q80Z29
-
-
-
r
nictoinamide + alpha-D-5-phosphoribosyl 1-diphosphate
nicotinamide mononucleotide + diphosphate
show the reaction diagram
-
whole cell lysates incubated with 5 microM [carbonyl-14C]nicotinamide + 0.5 mM alpha-D-5-phosphoribosyl-1-diphosphate at pH 8.8
produced NMN quantified by scintillation counting
-
r
phospho-NAMPT + H2O
NAMPT + phosphate
show the reaction diagram
-
DELTAG = -9.2 kcal/mol
-
-
?
ATP + NAMPT
ADP + phospho-NAMPT
show the reaction diagram
-
autophosphorylation (DELTAG = 1.9 kcal/mol, Keq = 0.047) is unfavourable and appears as initial burst of ADP generation
at 2.5 mM ATP and 46 microM NAMPT 77% of NAMPT is phosphorylated (proposed: His247 phospho-NAMPT), hydrolysis of phospho-NAMPT at 0.8/min, active phosphorylated NAMPT species neither isolated nor characterized yet
-
r
additional information
?
-
-
FK866, a highly specific noncompetitive inhibitor of nicotinamide phosphoribosyltransferase, represents a novel mechanism for induction of tumor cell apoptosis in tissues with a high demand for NAD+
-
-
-
additional information
?
-
Q99KQ4
pathway reconstitution in vitro using recombinant enzymes, overview
-
-
-
additional information
?
-
-
NMPRTase is a crucial enzyme in the salvage pathway of NAD+ biosynthesis and has important functions in regulating NAD+ levels in cells undergoing substantial NAD+ turnover
-
-
-
additional information
?
-
-
rate-limiting enzyme for NAD+ salvage from nicotinamide. Replicative senescence of smooth muscle cells is preceded by a marked decline in the expression and activity of Nampt. Nampt is a longevity protein that can add stress-resistant life to human smooth muscle cells by optimizing SIRT1-mediated p53 degradation
-
-
-
additional information
?
-
Q80Z29
visfatin mimics insulin signaling by binding to the insulin receptor with an affinity similar to that of insulin and does not share the binding site with insulin on the insulin receptor
-
-
-
additional information
?
-
-
facultative ATP hydrolysis activity, not tightly coupled to NMN synthesis
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
-
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
-
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
-
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
-
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
-
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
-
-
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
first step in NAD synthesis from nicotinamide
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
pre-B-cell colony enhancing factor
-
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
-
key enzyme in the regulation of NAD+ biosynthesis from the natural precursor nicotinamide
-
-
?
nicotinamide + 5-phospho-alpha-D-ribose 1-diphosphate
nicotinamide D-ribonucleotide + diphosphate
show the reaction diagram
Q99KQ4
key enzyme in the regulation of NAD+ biosynthesis, regulates the silent regulator protein 2, i.e. Sir2, activity, the enzyme has a metabolic regulatory function by up- and downregulation of proteins, overview
-
-
?
nicotinamide + alpha-D-5-phosphoribosyl 1-diphosphate
nicotinamide mononucleotide + diphosphate
show the reaction diagram
-
-
-
-
r
nicotinamide + alpha-D-5-phosphoribosyl 1-diphosphate
nicotinamide mononucleotide + diphosphate
show the reaction diagram
Q99KQ4
nicotinamide clearance
NMN synthesis is rate-limiting step for NAD+ synthesis
-
r
nicotinamide + alpha-D-5-phosphoribosyl 1-diphosphate
nicotinamide mononucleotide + diphosphate
show the reaction diagram
-
nicotinamide clearance
NMN synthesis is rate-limiting step for NAD+ synthesis
-
r
nicotinamide + alpha-D-5-phosphoribosyl-1-diphosphate
nicotinamide mononucleotide + diphosphate
show the reaction diagram
-
-
-
-
r
nicotinamide + alpha-D-5-phosphoribosyl-1-diphosphate
nicotinamide mononucleotide + inorganic diphosphate
show the reaction diagram
Q80Z29
-
-
-
r
additional information
?
-
-
FK866, a highly specific noncompetitive inhibitor of nicotinamide phosphoribosyltransferase, represents a novel mechanism for induction of tumor cell apoptosis in tissues with a high demand for NAD+
-
-
-
additional information
?
-
-
NMPRTase is a crucial enzyme in the salvage pathway of NAD+ biosynthesis and has important functions in regulating NAD+ levels in cells undergoing substantial NAD+ turnover
-
-
-
additional information
?
-
-
rate-limiting enzyme for NAD+ salvage from nicotinamide. Replicative senescence of smooth muscle cells is preceded by a marked decline in the expression and activity of Nampt. Nampt is a longevity protein that can add stress-resistant life to human smooth muscle cells by optimizing SIRT1-mediated p53 degradation
-
-
-
additional information
?
-
Q80Z29
visfatin mimics insulin signaling by binding to the insulin receptor with an affinity similar to that of insulin and does not share the binding site with insulin on the insulin receptor
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
ATP
-
requirement
ATP
-
activation, up to 0.4 mM
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Mg2+
-
Km-value: 0.6 mM
Mg2+
-
requirement
Mg2+
-
activation, up to 10 mM
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
(4',5,7-trihydroxyflavone)-(3'->8)-(4',5,7-trihydroxyflavone)
-
-
(4',7-dimethoxy-5-hydroxyflavone)-(3'->8)-(4',7-dimethoxy-5-hydroxyflavone)
-
-
3-Acetylpyridine
-
weak, kinetics
3-acetylpyridine adenine dinucleotide
-
kinetics
5-Fluoronicotinamide
-
weak, kinetics
5P-DADMe-NMN
-
poor, competitive inhibitor of non-ATP and ATP coupled NMN synthesis with respect to PRPP and nicotinamide, 3-fold increased binding in presence of ATP
6-Aminonicotinamide
-
weak, kinetics
alpha-D-5-phosphoribosyl 1-diphosphate
-
inhibits ATPase activity at 1 mM concentration, nearly complete inhibition of ADP-ATP exchange at 100 microM
Alpha-NAD+
-
weak
APO866
-
also known as FK866, (E)-N-[4-(1-benzoylpiperidin-4-yl)butyl]-3-(pyridine-3-yl)-acrylamide, no impact on cell viability, dose-dependent reduction of levels of intracellular NAD in and secretion of TNFalpha, IL-1beta, IL-6 (pro-inflammatory cytokines) by monocyctes, monocyte-derived dendritic cells and total PBMCs upon stimulation of inflammatory response by 100 ng/ml lipopolysaccharide (LPS) or 5 microgram/ml Pansorbin (Staphylococcus aureus cells, SAC)
APO866
Q99KQ4
also known as FK866, (E)-N-[4-(1-benzoylpiperidin-4-yl)butyl]-3-(pyridine-3-yl)-acrylamide, 10 mg/kg, DBA/1 mice: beneficial effects on collagen-induced arthritis (CIA) due to impaired secretion of inflammatory cytokines (reduction of the mean arthritic score and number of affected paws from mice with CIA as well as decrease in local levels of IL-1beta and IL-6 but no impact on IL-10, IFN-gamma, CCL5 and IL-12p70 levels, decrease in inflammatory infiltrate and hyperplasia in knees and paws from mice with CIA) but not due to toxicity (no premature death of individuals, no impact on histology of liver, spleen, lung, gut, kidney, inguinal lymph nodes and brain, normal liver alanine aminotransferase levels, no enhanced apoptosis in arthritic paws) and not due to impact on anti-collagen immune response (similar anti-collagen IgG levels); also known as FK866, (E)-N-[4-(1-benzoylpiperidin-4-yl)butyl]-3-(pyridine-3-yl)-acrylamide, 10 mg/kg, isolated peritoneal exudate inflammatory cells (PEC) from C57BL/6 mice: time-dependent NAD depletion (lowest at 9 h, recovery after 14 h), addition of 10 mM NMN abolishes a reduction of intracellular NAD+ concentration as well as TNFalpha and IL-6 secretion (pro-inflammatory cytokines) in PECs induced by in vivo or in vitro stimulation of inflammatory response with 5 microgram/ml Pansorbin (Staphylococcus aureus cells, SAC) or 100 ng/ml lipopolysaccharide (LPS)
CHS-828
-
competitive inhibitor
FK-866
Q80Z29
the nicotinamide-binding site is important for inhibition by FK-866
FK-866
-
i.e. (2E)-N-[4-(1-benzoylpiperidin-4-yl)butyl]-3-phenylprop-2-enamide
FK-866
-
i.e. (2E)-N-[4-(1-benzoylpiperidin-4-yl)butyl]-3-phenylprop-2-enamide
FK866
-
a highly specific noncompetitive inhibitor the enzyme, represents a novel mechanism for induction of tumor cell apoptosis, counteracted by nicotinic acid and nicotinamide
FK866
-
Nampt antagonist, 10 nM, reduction of Nampt activity in SMCs to 22 +/-2% of baseline activity accompanied with shortened senescent-free survival; reducing Nampt activity with the antagonist FK866 induces premature senescence in smooth muscle cells
FK866
-
FK866 is bound in a tunnel at the interface of the NMPRTase dimer, the inhibitor can reduce cellular NAD+ levels and induce apoptosis in tumors
FK866
-
powerful, non-competitive inhibitor with and without ATP
gallotanine
-
i.e. (2R,3S,5R,6S)-tetrahydro-2H-pyran-2,3,4,5,6-pentaylpentakis(oxycarbonyl-5,6-dihydroxybenzene-3,1-diyl) pentakis(3,4,5-trihydroxybenzoate)
GMX1777
-
i.e. 1-[2-(2-(2-(2-methoxyethoxy)-ethoxy)ethoxy)-ethoxy-carbonyloxymethyl]-4-[N'-cyano-N''-(6-(4-chlorophenyl)-hexyl)-N-guanidino]pyridinium chloride, prodrug of GMX1778. A dose of 75 mg/kg administered over a 24 h intravenous infusion causes tumor regression in the IM-9 model, the SHP-77 model, and HCT-116 model. A 72 h continuous intravenous infusion is also effective in the IM-9 model, but is associated with smaller therapeutic index
GMX1777
-
i.e. 1-[2-(2-(2-(2-methoxyethoxy)-ethoxy)ethoxy)-ethoxy-carbonyloxymethyl]-4-[N'-cyano-N''-(6-(4-chlorophenyl)-hexyl)-N-guanidino]pyridinium chloride, prodrug of GMX1778. A dose of 75 mg/kg administered over a 24 h intravenous infusion produces GMX1778 steady-state plasma levels of about 1 microg/ml and causes nicotinamide dinucleotide levels to decrease significnantly in tumors. Nicotinic acid protects mice treated with a lethal dose of GMX1777
N-biphenyl-2-yl-8-(4-pyridin-3-yl-1H-1,2,3-triazol-1-yl)octanamide
-
displays an IC50 for cytotoxicity in vitro of 3.8 nM and an IC50 for NAD depletion of 3.0 nM. Compound induces autophagic cell death
-
NAD+
-
1 mM, 50%-90% inhibition in various tissues, at 0.2 mM not inhibitory in lung and liver; strong (thigh muscle)
NAD+
-
competitive inhibitor of non-ATP and ATP coupled NMN synthesis with respect to PRPP and nicotinamide
NADH
-
competitive inhibitor of non-ATP and ATP coupled NMN synthesis with respect to PRPP and nicotinamide
Nicotinamide
-
substrate inhibition
nicotinamide hypoxanthine dinucleotide
-
-
nicotinamide mononucleotide
-
kinetics
nicotinamide mononucleotide-H2
-
strong, kinetics
Nicotinamide riboside
-
kinetics
nicotinate adenine dinucleotide
-
weak
nicotinate adenine dinucleotide
-
not inhibitory
Thionicotinamide
-
weak, kinetics
Thionicotinamide adenine dinucleotide
-
-
TP201565
-
competitive inhibitor
hypoxanthine
-
phosphorolysis
additional information
-
no inhibition by NaF
-
additional information
-
nicotinic acid, thymine, 5-bromouracil
-
additional information
-
no impact on ADP-ATP exchange rate by 10 microM adenylate kinase inhibitor P(1), P(5)-di(adenosine-5-)pentaphosphate (Ap5A), NAMPT does not accommodate Ap5A
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
ATP
-
35-fold higher concentrations of NMN synthesised, impact on substrate affinities, coupling of ATP hydrolysis to NMN synthesis enhances catalytic efficiency, accelerates forward turnover and overcomes the unfavourable thermodynamic equilibrium
ATP
-
essential activator
diphosphate
-
enhanced ATPase activity
EDTA
-
50% activation, 1 mM
Nicotinamide
-
enhanced ATPase activity
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0038
-
5-phospho-alpha-D-ribose 1-diphosphate
-
-
0.00537
-
5-phospho-alpha-D-ribose 1-diphosphate
-
-
0.00051
-
5-phosphoribosyl 1-diphosphate
-
with 1 mM ATP, pH 7.5, 37C
0.0232
-
5-phosphoribosyl 1-diphosphate
-
without ATP, pH 7.5, 37C
0.44
-
ADP
-
+/-0.05 mM, ADP-ATP radiolabel exchange in absence of other reactants
0.00063
-
alpha-D-5-phosphoribosyl 1-diphosphate
-
+/-0.00003 mM, in presence of 2 mM ATP the Km is lowered 10-fold, calculated from apparent KM by extrapolation to correct for inhibition by NAD+ in the continuous fluorimetric assay, pH 7.5, 125 nM nicotinamide, 2.2 nM NAMPT
0.0072
-
alpha-D-5-phosphoribosyl 1-diphosphate
-
+/-0.0006 mM, pH 7.5, 1 microM nicotinamide, 88 nM NAMPT, HPLC-based quantification
0.0117
-
alpha-D-5-phosphoribosyl 1-diphosphate
-
+/-0.0008 mM, in presence of 2 mM phosphate, pH 7.5, 1 microM nicotinamide, 88 nM NAMPT, HPLC-based quantification
2.2
-
ATP
-
+/-0.2 mM, ADP-ATP radiolabel exchange in absence of other reactants
7.4
-
ATP
-
+/-1.5 mM, in presence of 2 mM phosphate, pH 7.5, 100 nM nicotinamide, 50 microM PRPP, 2.2 nM NAMPT
5e-06
-
Nicotinamide
-
+/-0.000002 mM, in presence of 2 mM ATP the Km is 150-fold lowered, calculated from apparent KM by extrapolation to correct for inhibition by NAD+ in the continuous fluorimetric assay, pH 7.5, 200 microM PRPP, 2.8 nM NAMPT
6.7e-05
-
Nicotinamide
-
-
0.000235
-
Nicotinamide
-
+/-0.000017 mM, in presence of 2 mM phosphate, pH 7.5, 100 microM PRPP, 88 nM NAMPT, HPLC-based quantification
0.000855
-
Nicotinamide
-
+/-0.000022 mM, pH 7.5, 100 microM PRPP, 88 nM NAMPT, HPLC-based quantification
0.001
-
Nicotinamide
-
in the presence of ATP
0.00122
-
Nicotinamide
-
with 1 mM ATP, pH 7.5, 37C
0.00124
-
Nicotinamide
-
-
0.00127
-
Nicotinamide
-
-
0.0016
-
Nicotinamide
-
-
0.00806
-
Nicotinamide
-
without ATP, pH 7.5, 37C
0.92
-
Nicotinamide
-
pH 7.4, 37C, recombinant enzyme
100
-
Nicotinamide
-
-
100
-
Nicotinamide
-
in the absence of ATP
0.18
-
phosphate
-
+/-0.02 mM, pH 7.5, 88 nM NAMPT, HPLC-based quantification
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0003
-
ATP
-
ATPase activity is decreased in simultaneous presence of 1 mM of substrates of forward and reverse NAMPT reaction
0.00183
-
ATP
-
ATPase (ATP hydrolysis) activity of the free NAMPT, 2 mM ATP
0.00617
-
ATP
-
ATPase activity is enhanced in presence of 1 mM nicotinamide, 2 mM ATP
0.015
-
ATP
-
ADP-ATP radiolabel exchange in absence of other reactants (without NMN synthesis), formation or hydrolysis of phospho-NAMPT may be rate limiting for overall NMN synthesis reaction in presence of ATP
0.0183
-
ATP
-
ATPase activity is enhanced in presence of 1 mM diphosphate, presence of PRPP and/or nicotinamide reverses stimulation by diphosphate, 2 mM ATP
0.0433
-
ATP
-
ATPase activity in presence of 1 mM imidodiphosphate (PNP) is higher than in presence of pyophosphate, 2 mM ATP
0.00137
-
Nicotinamide
-
+/-0.00002, KM/kcat = 1600 1/M*s, pH 7.5, 100 microM PRPP, 88 nM NAMPT
0.00142
-
Nicotinamide
-
+/-0.00002, in presence of 2 mM phosphate, KM/kcat = 6100 1/M*s, pH 7.5, 100 microM PRPP, 88 nM NAMPT
0.00767
-
Nicotinamide
-
+/-0.00150, in presence of 2 mM ATP the catalytic efficiency (KM/kcat = 1800000 1/M*s) is improved 1100-fold, pH 7.5, 100 microM PRPP, 88 nM NAMPT
0.02
-
Nicotinamide
-
pH 7.4, 37C, recombinant enzyme
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.00016
-
5P-DADMe-NMN
-
with respect to alpha-D-5-phosphoribosyl 1-diphosphate (PRPP), in presence of 2.5 mM ATP, KM/Ki = 4, 200 microM PRPP, 100 nM nicotinamide, 2.2 nM NAMPT, 15 min
0.00053
-
5P-DADMe-NMN
-
with respect to alpha-D-5-phosphoribosyl 1-diphosphate (PRPP), in absence of ATP, KM/Ki = 14, 100 microM PRPP, 1 microM nicotinamide, 88 nM NAMPT, 15 min
0.0025
-
5P-DADMe-NMN
-
with respect to nicotinamide, in presence of 2.5 mM ATP, KM/Ki <1, 200 microM PRPP, 100 nM nicotinamide, 2.2 nM NAMPT, 15 min
0.0003
-
FK866
-
free enzyme
0.00014
-
NAD+
-
with respect to alpha-D-5-phosphoribosyl 1-diphosphate (PRPP), in presence of 2.5 mM ATP, KM/Ki = 5, 200 microM PRPP, 100 nM nicotinamide, 2.2 nM NAMPT, 15 min
0.00022
-
NADH
-
with respect to alpha-D-5-phosphoribosyl 1-diphosphate (PRPP), in presence of 2.5 mM ATP, KM/Ki = 3, 200 microM PRPP, 100 nM nicotinamide, 2.2 nM NAMPT, 15 min
0.0032
-
NADH
-
with respect to nicotinamide, in presence of 2.5 mM ATP, KM/Ki <1, 200 microM PRPP, 100 nM nicotinamide, 2.2 nM NAMPT, 15 min
0.00025
-
Nicotinamide
-
NMN synthesis
0.003
-
Nicotinamide
-
NMN synthesis, in presence of 5 microM NAD+
0.0071
-
5P-DADMe-NMN
-
with respect to nicotinamide, in absence of ATP, KM/Ki <1, 100 microM PRPP, 1 microM nicotinamide, 88 nM NAMPT, 15 min
additional information
-
ATP
-
> 4 mM, inhibition of NMN synthesis by phosphorylated NAMPT
additional information
-
FK866
-
10 +/-1 pM, with respect to alpha-D-5-phosphoribosyl 1-diphosphate (PRPP), in presence of 2.5 mM ATP, KM/Ki = 63000, 200 microM PRPP, 100 nM nicotinamide, 2.2 nM NAMPT, 15 min; 150 +/-18 pM, with respect to nicotinamide, in presence of 2.5 mM ATP, KM/Ki = 33, 200 microM PRPP, 100 nM nicotinamide, 2.2 nM NAMPT, 15 min; 5000 +/-500 pM, with respect to nicotinamide, in absence of ATP, KM/Ki = 171, 100 microM PRPP, 1 microM nicotinamide, 88 nM NAMPT, 15 min; 730 +/-75 pM, with respect to alpha-D-5-phosphoribosyl 1-diphosphate (PRPP), in absence of ATP, KM/Ki = 9863, 100 microM PRPP, 1 microM nicotinamide, 88 nM NAMPT, 15 min
0.0004
-
FK866
-
enzyme-substrate complex
additional information
-
NAD+
-
> 100 microM, with respect to nicotinamide, in absence of ATP
0.0021
-
NAD+
-
with respect to nicotinamide, in presence of 2.5 mM ATP, KM/Ki <1, 200 microM PRPP, 100 nM nicotinamide, 2.2 nM NAMPT, 15 min
additional information
-
NADH
-
> 100 microM, with respect to nicotinamide, in absence of ATP
IC50 VALUE [mM]
IC50 VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0066
-
(4',5,7-trihydroxyflavone)-(3'->8)-(4',5,7-trihydroxyflavone)
-
pH 7.5, 37C
0.0175
-
(4',7-dimethoxy-5-hydroxyflavone)-(3'->8)-(4',7-dimethoxy-5-hydroxyflavone)
-
pH 7.5, 37C
9e-07
-
FK-866
-
i.e. (2E)-N-[4-(1-benzoylpiperidin-4-yl)butyl]-3-phenylprop-2-enamide, pH 7.5, 37C
0.00011
-
FK-866
-
wild type protein, pH 8.5, 37C
0.008585
-
FK-866
-
H191R mutant protein, pH 8.5, 37C
0.0013
-
gallotanine
-
i.e. (2R,3S,5R,6S)-tetrahydro-2H-pyran-2,3,4,5,6-pentaylpentakis(oxycarbonyl-5,6-dihydroxybenzene-3,1-diyl) pentakis(3,4,5-trihydroxybenzoate), pH 7.5, 37C
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.000225
-
-
-
0.00213
-
-
at 0.001 mM 5-phosphoribosyl 1-diphosphate, without ATP, pH 7.5, 37C
0.0259
-
-
at 0.050 mM 5-phosphoribosyl 1-diphosphate, without ATP, pH 7.5, 37C; substrate 5-phosphoribosyl 1-diphosphate, without ATP, pH 7.5, 37C
0.0393
-
-
substrate nicotinamide, without ATP, pH 7.5, 37C
0.0397
-
-
substrate nicotinamide, with 1 mM ATP, pH 7.5, 37C
0.0481
-
-
at 0.050 mM 5-phosphoribosyl 1-diphosphate, with 1 mM ATP, pH 7.5, 37C
0.0704
-
-
at 0.001 mM 5-phosphoribosyl 1-diphosphate, with 1 mM ATP, pH 7.5, 37C
0.0741
-
-
substrate 5-phosphoribosyl 1-diphosphate, with 1 mM ATP, pH 7.5, 37C
additional information
-
-
activity per g hemoglobin
additional information
-
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.8
7.6
-
-
7.4
-
-
assay at
7.5
-
-
for both, NMN synthesis and ATP hydrolysis activity
8.5
9
-
assay at pH 8.8. where NAD glycohydrolase is not active
8.8
-
-
assay at
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.1
8.5
-
about half-maximal activity at pH 6.1 and pH 8.5
6.5
8.5
-
preincubation of NAMPT for 1 h at 25 C and various pH values in presence of 1 microM nicotinamide, 1 mM ATP, 30 nM NAMPT (NAMPT reaction started with 200 microM alpha-D-5-phosphoribosyl 1-diphosphate) or 2 mM ATP, 0.75 microM NAMPT (ATP hydrolysis)
7.8
9.2
-
about half-maximal activity at pH 7.8 and about 80% of maximal activity at pH 9.2
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
35
-
-
assay at
37
-
-
assay at
37
-
-
assay at
37
-
-
assay at
37
-
-
assay at
37
-
-
assay at
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.69
-
-
theoretic value
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
primary SMC culture, Nampt expression declines as SMC approach senescence as detected by immunoblotting using rabbit-polyclonal anti-human Nampt/Pre-B-cell colony-enhancing factor antibody; primary SMC culture stably expressing Nampt, 2.1 +/-0.3 additional population doublings constituting 34 +/-4% lifespan prolongation
Manually annotated by BRENDA team
Q80Z29
similar mRNA expression levels in (fa/fa) Zucker rats (obese) compared to lean Zucker rats (control) as well as in cafeteria diet-fed Wistar rats (diet-induced obesity) compared to standard chow diet-fed Wistar rats (control)
Manually annotated by BRENDA team
Q99KQ4
positive staining in blood vessels from arthritic joints as revealed by immunohistochemistry using rat monoclonal anti-murine NAMPT antibody
Manually annotated by BRENDA team
Q99KQ4
some positive staining in cells of both normal and arthritic joints as revealed by immunohistochemistry using rat monoclonal anti-murine NAMPT antibody
Manually annotated by BRENDA team
-
diploid, GM 1362, from patient with Lesch-Nyhan disease
Manually annotated by BRENDA team
-
enzyme and Sir2 expression levels
Manually annotated by BRENDA team
Q80Z29
inguinal white adipose tissue
Manually annotated by BRENDA team
-
liver carcinoma cell line
Manually annotated by BRENDA team
-
release of NAMPT enzyme into the cell culture supernatant occurs constitutively. HepG2 lysates and supernatants primarily contain the dimeric form of NAMPT which exhibit similar in vitro specific enzymatic activity
Manually annotated by BRENDA team
-
release of NAMPT enzyme into the cell culture supernatant occurs constitutively. In primary hepatocytes, secretion within 24 h is far higher than the cellular content, but is neither influenced by inhibitors of secretion nor by glucose, insulin or TNFalpha. In primary human hepatocytes, secreted NAMPT is less active than intracellular enzyme
Manually annotated by BRENDA team
-
SMC clonal line, Nampt expression declines as SMC approach senescence as detected by immunoblotting using rabbit-polyclonal anti-human Nampt/Pre-B-cell colony-enhancing factor antibody, 14 +/-3% of basal Nampt activity in presenescent SMC, direct relationship between Nampt activity and number of replication cycles preceeding senescence; SMC clonal line stably expressing Nampt, 7.1 +/-3.1 fold higher Nampt activity, 6.3 +/-0.3 additional population doublings constituting 71 +/-7% lifespan prolongation, 19 +/-2% reduction in number of senescent cells at passage 37 compared to control HITC6 cells is dependent on SIRT1 activity and associated with increased p53 degradation and protection against oxidative cell damage as response to 150 microM hydrogen peroxide
Manually annotated by BRENDA team
Q99KQ4
massive expression in synoviocytes of the synovial lining layer, sub-intimal synovium and pannus in joints from mice with type II collagen induced arthritis (CIA) compared to non-arthritic naive DBA/1 mice as revealed by immunohistochemistry using rat monoclonal anti-murine NAMPT antibody, some positive staining in chondrocytes from normal and arthritic joints and in blood vessels and inflammatory cells from arthritic joints
Manually annotated by BRENDA team
-
leukemia cell line
Manually annotated by BRENDA team
-
isolated from total PBMCs from blood from healthy individuals
Manually annotated by BRENDA team
Q99KQ4
elevated levels in mice with type II collagen induced arthritis (CIA) compared to non-arthritic naive DBA/1 mice as revealed by ELISA using a mouse visfatin/PBEF ELISA kit
Manually annotated by BRENDA team
-
PBMC from blood from healthy individuals
Manually annotated by BRENDA team
Q99KQ4
(PEC) isolated from naive C57BL/6 mice of from lipopolysaccharide-treated mice
Manually annotated by BRENDA team
Q99KQ4
elevated levels upon type II collagen induced arthritis (CIA) compared to non-arthritic naive DBA/1 mice as revealed by ELISA using a mouse visfatin/PBEF ELISA kit
Manually annotated by BRENDA team
-
from individual with Hutchison-Gilford progeria syndrome associated with premature atherosclerosis, cell lifespan extension upon Nampt overexpression
Manually annotated by BRENDA team
Q80Z29
similar mRNA expression levels in mesenteric, retroperitoneal and epididymal WAT depots of (fa/fa) Zucker rats (obese) compared to lean Zucker rats (control)
Manually annotated by BRENDA team
Q80Z29
lean Wistar rats: mRNA expression levels are maximal in mesenteric WAT (2.01 +/-0.08) followed by inguinal (1.23 +/-0.15), epididymal (0.88 +/-0.13), and retroperitoneal (0.79 +/-0.07) WAT; lean Zucker rats: mRNA expression levels are similar in subcutaneous (inguinal, 1.18 +/-0.33) and visceral (mesenteric (0.99 +/-0.15), retroperitoneal (0.56 +/-0.10), epididymal (1.17 +/-0.14)) WAT; mesenteric, significantly decreased mRNA expression level in cafeteria diet-fed Wistar rats (diet-induced obesity) compared to standard chow diet-fed Wistar rats (control); significantly reduced mRNA expression level in (fa/fa) Zucker rats (obese) compared to lean Zucker rats (control) and in cafeteria diet-fed Wistar rats (diet-induced obesity) compared to standard chow diet-fed Wistar rats (control)
Manually annotated by BRENDA team
-
from blood from healthy individuals
Manually annotated by BRENDA team
additional information
-
tissue distribution
Manually annotated by BRENDA team
additional information
-
ubiquitous in all tissues
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
55000
-
-
gel filtration, part of the enzyme in supernatant of HepG2 cells
120000
-
-
gel filtration, most of the enzyme in supernatant of HepG2 cells
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 55600, deduced from gene sequence
?
-
x * 52000, immunoblot analysis
dimer
-
according to sedimentation-equilibrium measurements following ultracentrifugation of the protein solution
dimer
-
2 * 55000, SDS-PAGE, plus some monomer, supernatant of HepG2 cells
homodimer
-
two molecules in the asymmetric unit of the crystal according to Matthews coefficient Vm = 2.4 A/Da, corresponding to a solvent content of 48.5%
homodimer
-
2 * 55520, theoretic value
monomer
-
1 * 55000, SDS-PAGE, plus some dimer, supernatant of HepG2 cells
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
phosphoprotein
-
active phosphorylated NAMPT species neither isolated nor characterized
phosphoprotein
-
autophosphorylation can increase the activity
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
apo enzyme, diffraction of third crystals at 2 A resolution at -174C, crystals belong to space group P2(1) with unit-cell parameters a = 60.56 A, b = 106.40 A, c =82.78 A, beta = 96.50, technique: 1 microlitre of 10 mg/ml protein solution to 1 microlitre reservoir solution (38% pentaerytritol propoxylate (5/4 PO/OH) and 200 mM KCl pH 9) one week at 4C, generation of first crystals by hanging-drop vapour diffusion (Index HT screening kit), generation of second crystals by microseeding: reservoir solution contained crushed first crystals, generation of third crystals by Repeated Seeding Technique: reservoir solution contained crushed second crystals
-
crystal structures at up to 2.1 A resolution of NMPRTase, alone and in complex with the reaction product nicotinamide mononucleotide or the inhibitor FK866. Crystals of the selenomethionyl free enzyme of human NMPRTase (F132M I151M double mutant) are grown with the sitting drop vapor diffusion method at 22C. Crystals of wild-type human NMPRTase in complex with NMN are obtained at 22C by sitting drop vapor diffusion
-
crystal structures of the enzyme in the free form and bound to nicotinamide and 5-phospho-alpha-D-ribose 1-diphosphate at the resolution of 2.0 A to 2.2 A are essentially identical to that of the complex with nicotinamide mononucleotide, except for some variations that can facilitate the substitution reaction by fixing the nucleophile and the leaving group for the requisite inversion of configuration at the C1-carbon of the ribose ring. In the active site near the C1-atom of the bound 5-phospho-alpha-D-ribose 1-diphosphate or nicotinamide mononucleotide, there is neither negatively charged group nor waterproof environment necessary to support the feasibility of a ribo-oxocarbocation intermediate inherent in the SN1 mechanism
-
crystallization of recombinant human NMPRTase in the free form, sitting-drop vapour-diffusion method, hanging-drop vapour-diffusion method, crystal quality is improved by successive use of the microseeding technique. The resultant crystals diffract to 2.0 A resolution. These crystals belongs to space group P2(1), with unit-cell parameters a = 60.56, b = 106.40, c = 82.78 A
-
crystal structures at up to 2.1 A resolution of NMPRTase, alone and in complex with the reaction product nicotinamide mononucleotide or the inhibitor FK866. Crystals of wild-type murine NMPRTase free enzyme are obtained by sitting drop vapor diffusion at 4C
-
hanging drop vapor diffusion method, crystallization of apoenzyme and complex with either nicotinamide mononucleotide or the NAmPRTase inhibitor FK-866
Q80Z29
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.5
8.5
-
preincubation of 30 nM NAMPT for 1 h at 25 C and various pH values, NMN synthesis: reaction started with 200 microM alpha-D-5-phosphoribosyl 1-diphosphate in presence of 1 microM nicotinamide, 1 mM ATP
7
10
-
stable
8.8
-
-
rapid inactivation below
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
65
-
-
10 min, stable, heat treatment during purification
70
-
-
5 min, stable, heat treatment during purification
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
freeze-thawing inactivates
-
bovine serum albumin, glutathione, cysteine, DTT, EDTA, cystine or H2O2 does not stabilize during storage
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-70C, in crude erythrocyte lysates, at least 30 days
-
-80C, crude, 4-6 months
-
4C, crude, inactivation overnight
-
-20C to +20C, about 70% loss of activity overnight
-
storage at room temperature, 4C, or frozen, unstable upon
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
immobilized metal ion affinity chromatography (Ni2+)
-
lysis of bacteria by French press, elution from Ni-NTA column by 100 and 200 mM imidazole, concentration and desalting, elution from HiLoad Superdex 200GP 26/60 column with 100 mM HEPES pH 7.5, 100 mM NaCl, 10 mM beta-mercaptoethanol, concentrated to 50-75 mg/ml, kept frozen
-
lysis of bacteria by ultrasonification in presence of 1 mM phenylmethylsulfonylfluorid (PMSF) and 1 mM dithiothreitol (DTT), ultracentrifugation, supernatant applied to glutathione Sepharose 4B beads and washed, cleavage of glutathione S-transferase (GST) tag by PreScission Protease, purification by gel-filtration chromatography in Tris-HCl pH 8.5, 150 mM NaCl, 1 mM DTT, concentration to 10 mg/ml by ultrafiltration on Microcon YM-10, stored at 4C
-
partial
-
partially frim K-562 leukemic cells
-
immobilized metal ion affinity chromatography
-
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
significant homology to human pre-B-cell colony enhancing factor
-
a retroviral gene delivery system is used to generate human cells stably overexpressing Nampt. Introducing the Nampt gene into aging human SMCs delays senescence and substantially lengthened cell lifespan, together with enhanced resistance to oxidative stress; in pQCXIP-IHRES-PURO for retroviral delivery and stable overexpression in Phoenix-amphotropic retrovirus packaging cell line
-
cDNA into pGEX6p1 for expression in Escherichia coli BL21 (DE) as glutathione S-transferase (GST) tagged fusion protein
-
chemically synthesised DNA (DNA 2.0) in pBAD DEST 49 for expression in Escherichia coli BL21 (DE3)-pLys
-
expressed in Escherichia coli Rosetta 2 (DE3)
-
expression in Escherichia coli
-
expression in Escherichia coli BL21
-
DNA sequence determination and analysis, phylogenetic analysis, expression of His-tagged enzyme and of enzyme fused to Sir2 in Escherichia coli strain BL21(DE3)
-
expressed in Escherichia coli BL21-CodonPlus(DE3)-RIL
-
expression in Escherichia coli
-
pre-B-cell colony enhancing factor
-
expression in Escherichia coli
Q80Z29
EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
increased during macrophage differentiation
-
expression of Nampt in the heart is significantly decreased by ischemia, ischemia/reperfusion and pressure overload. Downregulation of Nampt increases caspase 3 cleavage, cytochrome c release, and TUNEL-positive cells, which are inhibited in the presence of Bcl-xL, but do not increase hairpin 2positive cells, suggesting that endogenous Nampt negatively regulates apoptosis but not necrosis. Downregulation of Nampt also impairs autophagic flux
-
upon aging: enzyme level increases in serum but decreases in brain, decreases in cortex and hippocampus but remains unchanged in cerebellum and striatum in brain, and increases in microglia but likely decreases in neuron
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
DELTA D93
-
naturally occuring mutation in CHS-828 or APO866 resistant cell lines, about 20% of wild type aczivity
H191R
-
activity reduced by about 50%
H191R/K342R
-
naturally occuring mutation in APO866 resistant cell line
H191R/K342R/Q388R
-
naturally occuring mutation in TP201565 resistant cell line
H247A
-
active site residue, no detectable activity
H247E
-
active site residue, low activity
K342R
-
naturally occuring mutation, activity reduced by about 50%
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
drug development
-
inhibitor-soaking experiments
medicine
-
longevity enzyme for human SMCs
medicine
-
a dose of 75 mg/kg GMX1777, i.e. 1-[2-(2-(2-(2-methoxyethoxy)-ethoxy)ethoxy)-ethoxy-carbonyloxymethyl]-4-[N'-cyano-N''-(6-(4-chlorophenyl)-hexyl)-N-guanidino]pyridinium chloride, prodrug of GMX1778, administered over a 24 h intravenous infusion causes tumor regression in the IM-9 model, the SHP-77 model, and HCT-116 model. A 72 h continuous intravenous infusion is also effective in the IM-9 model, but is associated with smaller therapeutic index
pharmacology
-
enzyme is a potential target for development of anticancer drugs
pharmacology
-
NAMPT inhibition might have therapeutic efficacy in immune-mediated inflammatory diseases through impact on inflammatory cytokine secretion by leukocytes
medicine
-
a dose of 75 mg/kg GMX1777, i.e. 1-[2-(2-(2-(2-methoxyethoxy)-ethoxy)ethoxy)-ethoxy-carbonyloxymethyl]-4-[N'-cyano-N''-(6-(4-chlorophenyl)-hexyl)-N-guanidino]pyridinium chloride, prodrug of GMX1778, administered over a 24 h intravenous infusion produces GMX1778 steady-state plasma levels of about 1 microg/ml and causes nicotinamide dinucleotide levels to decrease significnantly in tumors. Nicotinic acid protects mice treated with a lethal dose of GMX1777