Information on EC 3.2.2.5 - NAD+ glycohydrolase

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

EC NUMBER
COMMENTARY hide
3.2.2.5
-
RECOMMENDED NAME
GeneOntology No.
NAD+ glycohydrolase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
NAD+ + H2O = ADP-D-ribose + nicotinamide
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of N-glycosyl bond
-
-
-
-
P-O bond cleavage
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Metabolic pathways
-
-
Nicotinate and nicotinamide metabolism
-
-
NAD metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
NAD+ glycohydrolase
This enzyme catalyses the hydrolysis of NAD+, without associated ADP-ribosyl cyclase activity (unlike the metazoan enzyme EC 3.2.2.6, bifunctional ADP-ribosyl cyclase/cyclic ADP-ribose hydrolase). The enzyme from Group A streptococci has been implicated in the pathogenesis of diseases such as streptococcal toxic shock-like syndrome (STSS) and necrotizing fasciitis. The enzyme from the venom of the snake Agkistrodon acutus also catalyses EC 3.6.1.5, apyrase [3].
CAS REGISTRY NUMBER
COMMENTARY hide
9032-65-9
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
NAD+ glycohydrolase activity of C3 transferase
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
M serotype 6
-
-
Manually annotated by BRENDA team
diverse strains, e.g. strain SF370, of low and high NADase activity, as isolates taken from streptococcal toxic shock-like syndrome, STSS, patients, gene nga
-
-
Manually annotated by BRENDA team
group A, e.g. virulent M-type 3 strain
-
-
Manually annotated by BRENDA team
group A, e.g. virulent M-type 3 strain
-
-
Manually annotated by BRENDA team
NRRL B-2682
-
-
Manually annotated by BRENDA team
NRRL B-2682
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1,N6-etheno NAD+ + H2O
epsilon-ADP-ribose + nicotinamide + H+
show the reaction diagram
-
-
-
-
?
1,N6-etheno-NAD+ + H2O
?
show the reaction diagram
2-fluoro-NAD+ + H2O
2-fluoro-ADP-ribose + nicotinamide
show the reaction diagram
-
in presence of methanol, formation of methanolysis product beta-1''-O-methyl 2-fluoro-ADP-ribose, with a preference for methanolysis over hydrolysis of 100:1
-
-
?
3-acetylpyridine + H2O
?
show the reaction diagram
-
-
-
-
?
3-acetylpyridine adenine dinucleotide + H2O
?
show the reaction diagram
3-acetylpyridine hypoxanthine dinucleotide + H2O
?
show the reaction diagram
3-aminopyridine + H2O
?
show the reaction diagram
-
-
-
-
?
3-carboxyhydrazide adenine dinucleotide + H2O
?
show the reaction diagram
-
-
-
-
?
3-formylpyridine adenine dinucleotide + H2O
?
show the reaction diagram
-
-
-
-
?
3-methylpyridine + H2O
?
show the reaction diagram
-
-
-
-
?
3-pyridylacetonitrile + H2O
?
show the reaction diagram
-
-
-
-
?
3-pyridylcarbinol + H2O
?
show the reaction diagram
-
-
-
-
?
alpha-NAD+ + H2O
ADP-ribose + nicotinamide
show the reaction diagram
-
-
-
-
?
beta-NAD+ + H2O
ADP-ribose + nicotinamide
show the reaction diagram
beta-NAD+ + H2O
ADP-ribose + nicotinamide + H+
show the reaction diagram
-
-
-
-
?
beta-nicotine guanine dinucleotide + H2O
?
show the reaction diagram
-
-
-
-
?
dNAD+ + H2O
?
show the reaction diagram
methylnicotinate adenine dinucleotide + H2O
?
show the reaction diagram
-
-
-
-
?
NAD+ + H2O
adenosine + nicotinamide
show the reaction diagram
-
solubilized enzyme form sNADase, unusual cleavage reaction, no hydrolysis of the labile quarternary nicotinamide-ribose pyridinium linkage
-
-
?
NAD+ + H2O
ADP-D-ribose + nicotinamide
show the reaction diagram
NAD+ + H2O
ADP-ribose + nicotinamide
show the reaction diagram
NAD+ + H2O
ADP-ribose + nicotinamide + H+
show the reaction diagram
-
-
-
-
?
NAD+ + H2O
ADPribose + nicotinamide
show the reaction diagram
NAD+ + H2O
cADPribose + nicotinamide
show the reaction diagram
NADP+ + H2O
phospho-ADPribose + nicotinamide
show the reaction diagram
nicotinamide ethenoadenine dinucleotide + H2O
?
show the reaction diagram
nicotinamide guanine dinucleotide + H2O
3',5'-cyclic GDP-ribose + GDP-ribose
show the reaction diagram
-
-
-
-
?
nicotinamide guanine dinucleotide + H2O
?
show the reaction diagram
nicotinamide hypoxanthine dinucleotide + H2O
?
show the reaction diagram
nicotinic acid adenine dinucleotide + H2O
?
show the reaction diagram
-
-
-
-
?
NMN + H2O
phosphoribose + nicotinamide
show the reaction diagram
pyridine + H2O
?
show the reaction diagram
-
-
-
-
?
pyridine-3-aldehyde adenine dinucleotide + H2O
?
show the reaction diagram
pyridine-3-aldehyde hypoxanthine dinucleotide + H2O
?
show the reaction diagram
thioNAD+ + H2O
?
show the reaction diagram
thioNADP+ + H2O
?
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
NAD+ + H2O
ADP-D-ribose + nicotinamide
show the reaction diagram
NAD+ + H2O
ADP-ribose + nicotinamide
show the reaction diagram
NAD+ + H2O
ADPribose + nicotinamide
show the reaction diagram
NAD+ + H2O
cADPribose + nicotinamide
show the reaction diagram
-
-
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
-
AA-NADase has one strong and two weak Co2+ binding sites; the enzyme has one strong and two weak Co2+ binding sites
Mn2+
-
AA-NADase has one Mn2+ binding site; the enzyme has one Mn2+ binding site
Ni2+
-
AA-NADase has two strong and six weak Ni2+ binding sites; the enzyme has two strong and six weak Ni2+ binding sites
Zinc
-
purified bifunctional enzyme has a ADP-ribosyl cyclase/NAD glycohydrolase ratio of 1/120. In situ cyclase/NAD glycohydrolase ratio measured in seminal plasma is 1/1. Physiological concentrations of zinc present in the seminal fluid, in the range of 0.6 to 4 mM, are responsible for the modulation of the cyclase/NAD glycohydrolase ratio
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2'-deoxy-2'-fluoroarabinoside adenine dinucleotide
-
-
2-Hydroxy-3,5-diiodo-benzoic acid
-
-
2-Hydroxy-5-iodo-benzoic acid
-
-
3-acetylpyridine
3-aminobenzamide
-
-
4-Carboxyhydrazide-pyridine-adenine dinucleotide
-
-
8-azido-carba-NAD+
-
-
-
8-azido-pseudocarba-NAD+
-
-
-
adenosine
-
-
adenosine 3',5'-monophosphate
ADP-(2-deoxy-2-fluoro-D-arabinose)
the NAD analogue forms a covalent adduct after nicotinamide cleavage, resulting in inhibition of the enzyme activity, binding structure, overview
ADP-(2-deoxy-2-fluoro-D-ribose)
the NAD analogue forms a covalent adduct after nicotinamide cleavage, resulting in inhibition of the enzyme activity, binding structure, overview
ADP-ribose
ADP-ribosylated protein
-
-
-
ADPribose
Alpha-NAD+
-
5 mM, 70% inhibition
ara-NAD+
-
-
-
b-series gangliosides
-
-
-
beta-NAD+
-
substrate inhibition, mediated by ADP-ribosylation, partially reversible by arginine or histidine, inhibits apoptosis in vivo
beta-NADP+
-
in vivo
carba-NAD+
-
-
Cu2+
-
the enzyme has two classes of Cu2+ binding sites, one activator site with high affinity and approximately six inhibitor sites with low affinity
cyanidin
deamino-NAD+
-
5 mM, almost complete inhibition
delphinidin
-
-
diosmetinidin
-
-
dioxane
-
more than 0.05 mM inhibit the activity
dithiothreitol
-
inhibits both NADase and ADPase activities through the reduction of Cu(II) to Cu(I) and the cleavage of disulfide-bonds in AA-NADase
fisetinidin
-
-
gangliosides
-
-
-
GD1a
-
exogenous ganglioside, slight inhibition, inhibition in vivo
GD1b
-
exogenous ganglioside, inhibition in vivo
GD3
-
exogenous ganglioside, inhibition in vivo
GDP-ribose
-
-
glutathione
-
inhibits both NADase and ADPase activities through the reduction of Cu(II) to Cu(I) and the cleavage of disulfide-bonds in AA-NADase
GM1b
-
exogenous ganglioside, slight inhibition, inhibition in vivo
GT1b
-
exogenous ganglioside, inhibition in vivo, needs to be incorporated into the cell to inhibit CD38, preferably cis interaction, i.e. CD38 and GT1b are located on the same cell, the tandem sialic acid residues linked to the internal galactose of the gangliotetraose core are crucial to the inhibition
hydroxylamine
inhibits the reaction between ADP-ribose and polypeptides
iodoacetamide
-
-
Isoniazid
kuromanin
-
-
L-ascorbate
-
inhibits AA-NADase on both NADase and ADPase activities through the reduction of Cu(II) in AA-NADase to Cu(I)
luteolinidin
-
-
m-aminophenylboronic acid
-
-
monoclonal antibody CS/2
-
inhibits the NAD+ glycohydolase activity of both the isolated extracellular domain of CD38 and cell-surface CD38, non-competitive. No effect on cyclase activity of enzyme
-
myricetin
-
-
N1-cyclic inosine diphosphate ribose
-
NADP+
-
5 mM, almost complete inhibition
nicotinamide
nicotinic acid
NMN
-
5 mM, 70% inhibition
pelargonidin
-
-
petunidin
-
-
Procion Blue HB
-
-
protein IFS
-
pseudocarba-NAD+
-
quercetagetin
-
-
quercetagetinidin
-
-
quercetin
-
-
rac-taxifolin
-
-
robinetin
-
-
salicylic acid
-
-
Streptococcal NAD glycohydrolase inhibitor
Q7CFI6
the ifs gene, or SpyM3_0129, encodes an endogenous competitive inhibitor for the enzyme. IFS is localized intracellularly and forms a complex with the enzyme. This intracellular complex must be dissociated during export through the cell envelope. The interface between SPNct and IFS is highly rich in water molecules, binding structure, overview
-
streptococcal NADase inhibitor
-
161 amino acids, MW 18800 Da, thermostable. Monomeric NADase and the streptococcal NADase inhibitor rapidly form in vitro a stable heterodimer complex in the ratio 1.1, resulting in complete suppression of the hydrolase activity. The inhibitor protein, coexpressed and complexed with NADase, may protect the producer cocci from exhausion of NAD
-
thioNAD+
tris(2-carboxyethyl)phosphine
-
TCEP, inhibits both NADase and ADPase activities through the reduction of Cu(II) to Cu(I) and the cleavage of disulfide-bonds in AA-NADase
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
dioxane
-
up to 0.05 mM dioxane produces the more active monomeric form of the NADase
guanosine 5'-O-(3-thiotriphosphate)
-
enzyme may be regulated by a G protein-dependent mechanism
monoclonal antibody clone 90
-
stimulates the NAD+ glycohydrolase activity of the isolated extracellular domain of CD38, little effect on cell-surface CD38. No effect on cyclase activity of enzyme
-
additional information
-
increase in activity ba Triton X-100 treatment due to solubilization from the cells
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.016
2-fluoro-NAD+
-
pH 7.4
20
3-acetylpyridine
-
-
0.006 - 0.15
3-acetylpyridine adenine dinucleotide
0.007
3-acetylpyridine hypoxanthine dinucleotide
-
-
3.2
3-aminopyridine
-
-
0.4
3-carboxyhydrazide adenine dinucleotide
-
spleen
6.5
3-methylpyridine
-
-
6.1
3-pyridylacetonitrile
-
-
7.9
3-Pyridylcarbinol
-
-
0.16
beta-nicotinamide guanine dinucleotide
-
-
-
0.1
dNAD+
0.009 - 0.014
epsilon-NAD+
0.5
methylnicotinate adenine dinucleotide
-
spleen
0.0046 - 2.14
NAD+
0.005 - 0.13
NADP+
0.02
Nicotinamide ethenoadenine dinucleotide
-
-
0.032
nicotinamide guanine dinucleotide
-
-
0.005 - 0.02
nicotinamide hypoxanthine dinucleotide
1.3
NMN
1.4
Pyridine
-
-
0.011 - 0.057
thioNAD+
0.01
thioNADP+
-
-
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.021 - 8390
NAD+
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.86
NAD+
Deinagkistrodon acutus
-
pH 7.4, 37C
7
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.6
ADP-ribose
-
pH 7.4, 37C, recombinant wild-type enzyme
1.6
DTT
-
pH 7.2, 37C, purified enzyme
0.33 - 0.6
nicotinamide
9
nicotinic acid
-
pH 7.2, 37C, purified enzyme
additional information
additional information
-
inhibition kinetics, overview
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0023
cyanidin
Schistosoma mansoni
-
at 37C in 10 mM potassium phosphate buffer, pH 7.4, containing 0.05% (w/v) emulphogen
0.006
delphinidin
Schistosoma mansoni
-
at 37C in 10 mM potassium phosphate buffer, pH 7.4, containing 0.05% (w/v) emulphogen
0.0099
diosmetinidin
Schistosoma mansoni
-
at 37C in 10 mM potassium phosphate buffer, pH 7.4, containing 0.05% (w/v) emulphogen
2.48
DTT
Deinagkistrodon acutus
-
pH 7.4, 37C, inhibition of the NADase activity
0.0064
fisetinidin
Schistosoma mansoni
-
at 37C in 10 mM potassium phosphate buffer, pH 7.4, containing 0.05% (w/v) emulphogen
6.11
glutathione
Deinagkistrodon acutus
-
pH 7.4, 37C, inhibition of the NADase activity
0.0082
kuromanin
Schistosoma mansoni
-
at 37C in 10 mM potassium phosphate buffer, pH 7.4, containing 0.05% (w/v) emulphogen
1.79
L-ascorbate
Deinagkistrodon acutus
-
pH 7.4, 37C, inhibition of the NADase activity
0.0084
luteolin
Schistosoma mansoni
-
at 37C in 10 mM potassium phosphate buffer, pH 7.4, containing 0.05% (w/v) emulphogen
0.0059
luteolinidin
Schistosoma mansoni
-
at 37C in 10 mM potassium phosphate buffer, pH 7.4, containing 0.05% (w/v) emulphogen
0.008
malvidin
Schistosoma mansoni
-
at 37C in 10 mM potassium phosphate buffer, pH 7.4, containing 0.05% (w/v) emulphogen
0.022
myricetin
Schistosoma mansoni
-
at 37C in 10 mM potassium phosphate buffer, pH 7.4, containing 0.05% (w/v) emulphogen
0.26
N1-cyclic inosine diphosphate ribose
Homo sapiens
P28907
pH 4.5, 20-24C
0.0044
pelargonidin
Schistosoma mansoni
-
at 37C in 10 mM potassium phosphate buffer, pH 7.4, containing 0.05% (w/v) emulphogen
0.0056
peonidin
Schistosoma mansoni
-
at 37C in 10 mM potassium phosphate buffer, pH 7.4, containing 0.05% (w/v) emulphogen
0.0378
petunidin
Schistosoma mansoni
-
at 37C in 10 mM potassium phosphate buffer, pH 7.4, containing 0.05% (w/v) emulphogen
0.0013
quercetagetin
Schistosoma mansoni
-
at 37C in 10 mM potassium phosphate buffer, pH 7.4, containing 0.05% (w/v) emulphogen
0.0061
quercetagetinidin
Schistosoma mansoni
-
at 37C in 10 mM potassium phosphate buffer, pH 7.4, containing 0.05% (w/v) emulphogen
0.0039
quercetin
Schistosoma mansoni
-
at 37C in 10 mM potassium phosphate buffer, pH 7.4, containing 0.05% (w/v) emulphogen
0.0195
rac-taxifolin
Schistosoma mansoni
-
at 37C in 10 mM potassium phosphate buffer, pH 7.4, containing 0.05% (w/v) emulphogen
0.0058
robinetin
Schistosoma mansoni
-
at 37C in 10 mM potassium phosphate buffer, pH 7.4, containing 0.05% (w/v) emulphogen
0.42
tris(2-carboxyethyl)phosphine
Deinagkistrodon acutus
-
pH 7.4, 37C, inhibition of the NADase activity
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.66
-
erythrocytes
12.8
-
purified enzyme
16.64
-
-
74.5
-
purified native enzyme from reticulocytes, pH 7.5, 37C
77.6
-
-
180
-
-
240
-
brain
420
-
spleen
511
-
liver plasma membrane
1316
-
liver, microsomal
7600
-
brain
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.3 - 7
-
nuclear membrane
6.5 - 7
6.5
-
nuclear envelope
6.6
-
purified enzyme
6.6 - 7.1
-
DNA associated NADase
6.7
-
plasma membrane
6.8
-
nucleus
7.5
-
assay at
7.6 - 8
-
chromatin NADase
8.2
-
erythrocytes
8.3
-
detergent solubilized NADase
8.5
-
steapsin treated enzyme
9
-
spleen
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5 - 9
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
32
-
assay at
36
-
steapsin treated enzyme
39
-
detergent solubilized NADase
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
lung epithelial cell line
Manually annotated by BRENDA team
-
TRAP(+) osteoclast formation is inhibitied by exogenous addition of enzyme or of ADPribose
Manually annotated by BRENDA team
-
enzyme level and ADP-ribosylation is significantly higher in patients and are correlated with tumour stage
Manually annotated by BRENDA team
-
leukemic cells, retinoic acid-treated
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
-
from bone marrow. Tumor necrosis factor alpha upregulates CD38. Induced CD38 expression enhances inflammatory gene expression by decreasing ERK1/2 phosphorylation and increasing NF kappaB activation. It negatively affects the expression of osteoclast markers. CD38 may reduce osteoclastogenesis and increase inflammatory gene induction by decreasing cellular histone deacetylase activity
Manually annotated by BRENDA team
-
CD38 is localized in or on sympathetic nerve terminals of this blood vessel
Manually annotated by BRENDA team
-
CD38 is localized in or on sympathetic nerve terminals of this blood vessel
Manually annotated by BRENDA team
-
sympathic nerve terminal of mesenteric artery or vein
Manually annotated by BRENDA team
-
bone marrow neutrophil
Manually annotated by BRENDA team
-
high enzyme levels in serum are associated with a poor prognosis in patients with colorectal cancer
Manually annotated by BRENDA team
-
enzyme is partially GPI-anchored, T-cell lacking ADP-ribosyltransferase activity
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
12000
-
gel filtration
27000
-
gel filtration
32000
x * 32000, recombinant enzyme HvnB, SDS-PAGE
32769
-
x * 40000, SDS-PAGE, x * 32769, sequence calculation
40000
-
x * 40000, SDS-PAGE, x * 32769, sequence calculation
48600
-
x * 48600, recombinant C-terminally HA-tagged and His6-tagged enzyme variant SPNJ4 from Escherichia coli strain TOP10, SDS-PAGE
62000
-
2 * 62000, SDS-PAGE
63000
-
gel filtration
83000
-
gel filtration and sucrose density gradient sedimentation
98000
-
gel filtration
100000
125000
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
proteolytic modification
-
NADase is synthesized as precursor with feeble activity. Activation by removing the carboxyl terminal region during or after secretion into culture medium
side-chain modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant wild-type and mutant enzymes, mixing of 0.001 ml of protein sample with 0.001 ml of precipitant solution containing 0.1 M imidazole, pH 7.5, and 12-24% PEG 4000, X-ray diffraction structure determination and analysis at 1.75-2.18 A resolution
-
CD38 in complex with ribosyl-2'-fluoro-deoxy-adenosine diphosphate or arabinosyl-2'-fluoro-deoxy-adenosine diphosphate ribose, soaking crystals in a solution containing either 5.2 mM inhibitor, and 100 mM MES, pH 6.0, 15% PEG 4000, and 30% glycerol, X-ray diffraction structure determination and analysis at 1.75-2.0 A resolution
wild-type and mutant E226Q in complex with cyclic ADP-ribose at 1.5 A resolution, with cyclic GDP-ribose at 1.68 A, and with NGD+ at 2.1A. Binding of cyclic ADP-ribose or cyclic GDP-ribose induce structural changes in the dipeptide E146D147 of 2.7 A. Resiudue E226 is critical in catalysis and in positioning of cyclic ADP-ribose; wild-type and mutant E226Q in complex with NAD+, NGD+, or GDP-ribose. The reaction intermediate is stabilized by polar interactions with the catalytic residue E226 rather than by a covalent linkage
-
wild-type and mutant E226Q in complex with inhibitor N1-cyclic inosine diphosphate ribose at 1.7 and 1.176 A resolution, respectively
C-terminal glycohydrolase domain of SPN (residues 191-451) in complex with IFS, vapor diffusion method, using 20%-25% (w/v) PEG 8K and 100 mM Tris-HCl (pH 8.0), at 22C
-
purified recombinant selenomethionine-labeled enzyme C-terminal domain (residues 193-451) in complex with the full-length inhibitor IFS, sitting drop vapor-diffusion method, mixing of 0.001 ml of protein solution with 0.001 ml of reservoir solution of 20% w/v tacsimate, pH 4.0, and 20% w/v PEG 3350, 22C, X-ray diffraction structure determination and analysis at 1.70 A resolution, single-wavelength anomalous diffraction, modelling
Q7CFI6
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5
-
5 min, 60C, retains of 55% of activity
137054
8.5
-
apruptly dropping of activity above
137059
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
24 h in 0.1%, w/v emulphogene 85% of activity is remained
47
-
90 min, no significant loss of activity
75
-
30 min, half of activity lost
80
-
1-2 min, complete inactivation
100
-
5 min, complete inactivation
additional information
-
stable to heat, activation of enzyme through boiling
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
chromatin NADase is sensitiv to Dnase
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
deoxycholate
-
50% inhibition at 0.05% w/v, fully reversible on dilution
dithiothreitol
-
1 mM, slight inactivation
emulphogene BC-720
-
no inhibitory effect
taurocholate
-
50% inhibition at 0.2% w/v, fully reversible on dilution
Triton X-100
-
no inhibitory effect
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-15C, potassium phosphate buffer, pH 7.4, 2 months
-
-20C, 6 months, phosphate buffer, 30%, w/v glycerol
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-20C, at least one month
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-20C, emulphogene, several months
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-20C, over 1 year
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-20C, several weeks
-
-80C, several months
-
4C, 5 days, loss of 50% of activity
-
4C, 7 days
-
4C, potassium phosphate buffer, pH 7.4, loss of 40% of activity in 4 weeks
-
4C, several months
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
15.6fold to homogeneity, rapid 3-step chromatographic procedure
-
Blue Sepharose column chromatography, gel filtration
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from seminal plasma
-
native enzyme
-
native enzyme 81fold from reticulocytes, by lipid extracction, affinity and hydrophobic interaction chromatography, and gel filtration
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native enzyme from venom
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native enzymes partially from culture supernatant, recombinant enzymes from Escherichia coli
partial
-
partially, solubilization from the membrane fraction, best with CHAPS or Triton X-100
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particulate pNADase from brain, solubilization to give soluble sNADase
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recombinant C-terminally HA-tagged and His6-tagged enzyme variant SPNJ4 from Escherichia coli strain TOP10
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recombinant His-tagged wild-type and E391Q mutant SPN from Escherichia coli
-
recombinant His-tagged wild-type and mutant enzymes from Pichia apstoris culture supernatant by nickel affinity chromatography
recombinant N-terminally His6-tagged enzyme from Escherichia coli strain Rosetta2 (DE3) by nickel affinity chromatography and gel filtration
Q7CFI6
Talon metal affinity column chromatography, Q-Sepharose column chromatography, and S-200 gel filtration
-
wild-type and mutant enzymes by anion exchange chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloning of inactive enzyme variant SPNH5 from strain HSC5 and of active variant SPNJ4, expression of C-terminally HA-tagged and His6-tagged variant SPNJ4 in Escherichia coli strain TOP10
-
co-expression of N-terminally His6-tagged enzyme with inhibitor IFS in Escherichia coli strain Rosetta2 (DE3)
Q7CFI6
DNA and amino acid sequence determination and analysis, genomic structure, recombinant expression of FLAG-tagged wild-type and mutant enzymes in HEK293 cells
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enzymatic domain of CD38; expression of extramembrane domain in Saccharomyces cerevisiae
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expressed in Pichia pastoris
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expression in Escherichia coli, FLAG-tag
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expression in THP-1 cells, selection of transfected cells by G418
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expression of extracellular region of CD38 in Drosophila S2 cells
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expression of wild-type and mutant enzymes in Pichia pastoris
gene hvnA, part of a four gene operon, and monocistronic gene hvnB, DNA and amino acid sequence determination and analysis, expression in Escherichia coli DH5alpha and BW23474
gene nga, cloning n Escherichia coli strain JM109, expression of wild-type and mutant enzymes
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genotyping, SPN has a different pattern of polymorphisms than Streptococcus pyogenes housekeeping genes. The ratio of nonsynonymous to synonymous nucleotide polymorphisms reveals different patterns of evolution for NADase-active and NADase-inactive strains. SPN amino acid sequence remains preserved despite loss of NADase activity, overview
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recombinant expression of His-tagged wild-type and E391Q mutant SPN in Escherichia coli as stable and soluble monomer in the periplasmic space
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recombinant expression of His-tagged wild-type and mutant enzymes in Pichia apstoris and secretion to the culture medium
the C-terminal glycohydrolase domain of SPN (residues 191-451) in complex with IFS is expressed in Escherichia coli strain DL41
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
clindamycin, CLI, induces the expression of Nga, and of the other two major toxins, streptolysin O and streptokinase, in the streptococcal toxic shock syndrome, in CLI-resistant isogenic covS mutant D2TYcovS strain, overview
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E179G
-
site-directed mutagenesis
F174G
-
site-directed mutagenesis, the point mutation F174G can turn ADP-ribosyl cyclase from a cyclase toward an NADase, overview
E226
-
crystallization data
E226Q
-
crystallization data
Q218A
-
site-directed mutagenesis
Q218D
-
site-directed mutagenesis, loss of NADase activity without increase in ADP-ribosyltransferase activity
Q218E
-
site-directed mutagenesis, loss of NADase activity and increase in ADP-ribosyltransferase activity
Q218A
-
site-directed mutagenesis
-
Q218D
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site-directed mutagenesis, loss of NADase activity without increase in ADP-ribosyltransferase activity
-
Q218E
-
site-directed mutagenesis, loss of NADase activity and increase in ADP-ribosyltransferase activity
-
D133A
site-directed mutagenesis
E124A
site-directed mutagenesis, the mutant is not inhibited by cyanidin in contrast to the wild-type enzyme
E202A
site-directed mutagenesis, the mutant is much less inhibited by cyanidin compared to the wild-type enzyme
H103A
site-directed mutagenesis, the mutant is less inhibited by cyanidin compared to the wild-type enzyme
H103F
site-directed mutagenesis
S169A
site-directed mutagenesis
W165A
site-directed mutagenesis, the mutant is inhibited by cyanidin like the wild-type enzyme
W165F
site-directed mutagenesis
D330G
-
mutation of the inactive enzyme variant SPNH5 causes only a slight increase in activity, but the mutation in SPNJ4 causes an increase in activity
DELTA41-140
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166% of wild-type activity, loss of translocation competence
DELTA41-190
-
221% of wild-type activity, loss of translocation competence
DELTA41-240
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3.5% of wild-type activity, loss of translocation competence
DELTA41-90
-
212% of wild-type activity, less stable than wild-type
E391Q
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a catalytically nearly inactive mutant, upon transformation, SPN is toxic to Saccharomyces cerevisiae, whereas a glycohydrolase-inactive SPN allows for viability
G330D
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mutant strain 188NADase(G330D), a derivative of 188 in which the chromosomal nga locus harbors a point mutation coding for amino acid substitution G330D in NADase. This mutation causes an enzymatically inactive NADase, associated with inactive enzyme in clinical isolates of Streptococcus pyogenes and in a deliberately constructed mutant
K289R
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swap at residue 289 of SPNH5 does not increase activity of the inactive enzyme variant but reduces actiivty of variant SPNJ4
K289R/D330G
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the mutation increases SPNH5 activity to the level of about 30% activity of active enzyme variant SPNJ4, but reduces activity of variant SPNJ4
L199I/Q253H/L280V
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swaps of the 3 N-terminal residues of the enzymatic domain of variant SPNJ4 do not affect activity
R289K/G330D/I374V
-
the mutations introduced into enzyme variant SPNJ4 result in an inactivation of the enzyme
R289K/I374V
-
swaps of the the 7th and 9th polymorphic residues of variant SPNJ4 result in less activation compared to mutation D330G alone
D330G
-
mutation of the inactive enzyme variant SPNH5 causes only a slight increase in activity, but the mutation in SPNJ4 causes an increase in activity
-
K289R
-
swap at residue 289 of SPNH5 does not increase activity of the inactive enzyme variant but reduces actiivty of variant SPNJ4
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K289R/D330G
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the mutation increases SPNH5 activity to the level of about 30% activity of active enzyme variant SPNJ4, but reduces activity of variant SPNJ4
-
L199I/Q253H/L280V
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swaps of the 3 N-terminal residues of the enzymatic domain of variant SPNJ4 do not affect activity
-
R289K/I374V
-
swaps of the the 7th and 9th polymorphic residues of variant SPNJ4 result in less activation compared to mutation D330G alone
-
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
-
simple, fast and reliable fluorescence-based assay method to evaluate the NAD+ glycohydrolase activity of cell-permeable C3 proteins
medicine