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Information on EC 6.3.5.1 - NAD+ synthase (glutamine-hydrolysing) and Organism(s) Mycobacterium tuberculosis and UniProt Accession P9WJJ3

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EC Tree
IUBMB Comments
NH3 can act instead of glutamine (cf. EC 6.3.1.5 NAD+ synthase).
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Select one or more organisms in this record: ?
This record set is specific for:
Mycobacterium tuberculosis
UNIPROT: P9WJJ3
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Word Map
The taxonomic range for the selected organisms is: Mycobacterium tuberculosis
The enzyme appears in selected viruses and cellular organisms
Synonyms
nade1, nitrogen-regulatory protein, glutamine-dependent nad synthetase, glutamine-dependent nad+ synthetase, mtunade, glutamine-dependent nadegln, nicotinamide adenine dinucleotide synthetase enzyme, gsp38, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glutamine-dependent NAD+ synthetase
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General stress protein 38
-
-
-
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GSP38
-
-
-
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NAD synthetase (glutamine)
-
-
-
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NAD(+) synthase [glutamine-hydrolyzing]
-
-
-
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NAD+ synthetase
-
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NAD+ synthetase (glutamine-hydrolysing)
-
-
-
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NadE-679
Nicotinamide adenine dinucleotide synthetase (glutamine)
-
-
-
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Nitrogen-regulatory protein
-
-
-
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Sporulation protein outB
-
-
-
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Synthetase, nicotinamide adenine dinucleotide (glutamine)
-
-
-
-
PATHWAY SOURCE
PATHWAYS
-
-, -, -, -, -, -, -, -
SYSTEMATIC NAME
IUBMB Comments
deamido-NAD+:L-glutamine amido-ligase (AMP-forming)
NH3 can act instead of glutamine (cf. EC 6.3.1.5 NAD+ synthase).
CAS REGISTRY NUMBER
COMMENTARY hide
37318-70-0
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + deamido-NAD+ + L-glutamine + H2O
AMP + diphosphate + NAD+ + L-glutamate
show the reaction diagram
-
-
-
?
ATP + deamido-NAD+ + L-Gln
AMP + diphosphate + NAD+ + L-Glu
show the reaction diagram
-
-
-
-
?
ATP + deamido-NAD+ + L-glutamine + H2O
AMP + diphosphate + NAD+ + L-glutamate
show the reaction diagram
ATP + deamido-NAD+ + NH3 + H2O
AMP + diphosphate + NAD+
show the reaction diagram
ATP + deamido-NAD+ + NH4+
AMP + diphosphate + NAD+
show the reaction diagram
-
-
-
-
?
ATP + deamido-NAD+ + NH4+ + H2O
AMP + diphosphate + NAD+
show the reaction diagram
-
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + deamido-NAD+ + L-glutamine + H2O
AMP + diphosphate + NAD+ + L-glutamate
show the reaction diagram
-
-
-
?
ATP + deamido-NAD+ + L-glutamine + H2O
AMP + diphosphate + NAD+ + L-glutamate
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
ATP
structure of the ATP-binding site at the mtuNadE synthetase domain, overview
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
adenosine 5'-[alpha,beta-methylene]triphosphate
i.e. AMP-CPP, nonhydrolysable analogue of ATP
nicotinic acid-adenine dinucleotide
i.e. NaAD+
3-[([7-[5-(benzyloxy)-2-(methoxycarbonyl)-1H-indol-1-yl]heptyl]oxy)carbonyl]-1-methylpyridinium
-
-
4-[([7-[5-(benzyloxy)-2-(methoxycarbonyl)-1H-indol-1-yl]heptyl]oxy)carbonyl]-N,N,N-trimethylanilinium
-
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4-[([8-[5-(benzyloxy)-1H-indol-1-yl]octyl]oxy)carbonyl]-N,N,N-trimethylanilinium
-
-
4-[2-([9-[5-(benzyloxy)-2-(methoxycarbonyl)-1H-indol-1-yl]nonyl]oxy)-2-oxoethyl]-N,N,N-trimethylanilinium
-
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.1 - 2
ATP
pH and temperature not specified in the publication
1.5
L-glutamine
pH and temperature not specified in the publication
0.12 - 1.4
ATP
0.13 - 2.9
deamido-NAD+
1.3 - 20
L-Gln
1.1 - 1.6
L-glutamine
0.089 - 15
NH3
1.6 - 27
NH4+
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.6 - 3
ATP
0.5 - 3.7
deamido-NAD+
0.012 - 3
L-Gln
0.0045 - 3.3
NH3
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.1 - 5
ATP
3.7 - 5.7
deamido-NAD+
0.0015 - 0.42
L-Glu
0.051 - 0.22
NH3
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.7
adenosine 5'-[alpha,beta-methylene]triphosphate
pH and temperature not specified in the publication
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0633
3-[([7-[5-(benzyloxy)-2-(methoxycarbonyl)-1H-indol-1-yl]heptyl]oxy)carbonyl]-1-methylpyridinium
Mycobacterium tuberculosis
-
pH 8.5, 37°C
0.0218
4-[([7-[5-(benzyloxy)-2-(methoxycarbonyl)-1H-indol-1-yl]heptyl]oxy)carbonyl]-N,N,N-trimethylanilinium
Mycobacterium tuberculosis
-
pH 8.5, 37°C
0.0248
4-[([8-[5-(benzyloxy)-1H-indol-1-yl]octyl]oxy)carbonyl]-N,N,N-trimethylanilinium
Mycobacterium tuberculosis
-
pH 8.5, 37°C
0.0578
4-[2-([9-[5-(benzyloxy)-2-(methoxycarbonyl)-1H-indol-1-yl]nonyl]oxy)-2-oxoethyl]-N,N,N-trimethylanilinium
Mycobacterium tuberculosis
-
pH 8.5, 37°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.033
-
recombinant NadE-738, nitrogen donor glutamine
0.09
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glutamine-dependent activity
0.1
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recombinant NadE-738, nitrogen donor NH4+
0.23
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NH3 dependent activity
0.44
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recombinant NadE-679, nitrogen donor glutamine
0.67
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recombinant NadE-679, nitrogen donor NH4+
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8
-
recombinant NadE-679, nitrogen donor NH4+
8.5
-
recombinant NadE-738, nitrogen donor NH4+, recombinant NadE-679, nitrogen donor glutamine
8.8
-
recombinant NadE-738, nitrogen donor glutamine
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
gene nadE
SwissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
most residues lining the ATP-binding site are conserved among the glutamine-dependent NAD+ synthetases, whereas residues not conserved, such as Leu399 andGly366, interact with the adenine ring and the adenylyl ribose with the backbone oxygen and nitrogen
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
600000
-
gel filtration, NadE-679
67000
-
8 * 67000, SDS-PAGE
98000
-
x * 98000, SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 98000, SDS-PAGE
octamer
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8 * 67000, SDS-PAGE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
wild-type mtuNadE in complex with 6-diazo-5-oxo-L-norleucine/NaAD+ and mutant enzymes, hanging drop vapour diffusion method, different combinations of the ligands 3 mM NaAD+, 4 mM ATP, 7 mM adenosine 5'-[alpha,beta-methylene]triphosphate, 20 mM L-glutamine, 6 mM NAD+, 10 mM AMP, 4 mM diphosphate, 100 mM L-glutamate, and 10 mM or 120 mM MgCl2 are used in the co-crystallization experiments, crystallization solution contains 1.2-1.6 M ammonium citrate tribasic, pH 6.5-8.0, with 5-15% v/v glycerol, 20°C, X-ray diffraction structure determination analysis at 2.0-2.85 A resolution, molecular replacement
native enzyme and selenomethionine-derivative, to 2.35 and 3.0 A resolution, respectively. Homooctameric subunit organization suggesting a tight dependence of catalysis on the quaternary structure, a 40 A intersubunit ammonia tunnel and structural elements that may be involved in the transfer of information between catalytic sites
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C176A
site-directed mutagenesis, inactive mutant, analysis of ligand binding structures
L486A
site-directed mutagenesis
L486F
site-directed mutagenesis
Y58A
site-directed mutagenesis
C176A
D656A
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strong decrease in catalytic efficiency
E52A
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complete loss of glutamine-dependent activity, retains 30% of its NH3-dependent activity
K121A
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complete loss of glutamine-dependent and NH3-dependent activity
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
mutant enzymes expressed in Escherichia coli
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recombinant NAD+ synthase, Ni2+ chelate resin affinity chromatography
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recombinant NadE-738 and NadE-679, Ni2+-nitrilotriacetic acid agarose, gel filtration
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
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expression of His-tagged NAD+ synthase in Escherichia coli
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most efficient expression of His-tagged NadE-738 and NadE-679 in Escherichia coli strain Origami
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Cantoni, R.; Branzoni, M.; Labo, M.; Rizzi, M.; Riccardi, G.
The MTCY428.08 gene of Mycobacterium tuberculosis codes for NAD+ synthetase
J. Bacteriol.
180
3218-3221
1998
Mycobacterium tuberculosis
Manually annotated by BRENDA team
Bellinzoni, M.; De Rossi, E.; Branzoni, M.; Milano, A.; Peverali, F.A.; Rizzi, M.; Riccardi, G.
Heterologous expression, purification, and enzymatic activity of Mycobacterium tuberculosis NAD(+) synthetase
Protein Expr. Purif.
25
547-557
2002
Mycobacterium tuberculosis
Manually annotated by BRENDA team
Bellinzoni, M.; Buroni, S.; Pasca, M.R.; Guglierame, P.; Arcesi, F.; De Rossi, E.; Riccardi, G.
Glutamine amidotransferase activity of NAD+ synthetase from Mycobacterium tuberculosis depends on an amino-terminal nitrilase domain
Res. Microbiol.
156
173-177
2005
Mycobacterium tuberculosis
Manually annotated by BRENDA team
Boshoff, H.I.; Xu, X.; Tahlan, K.; Dowd, C.S.; Pethe, K.; Camacho, L.R.; Park, T.H.; Yun, C.S.; Schnappinger, D.; Ehrt, S.; Williams, K.J.; Barry, C.E.
Biosynthesis and recycling of nicotinamide cofactors in Mycobacterium tuberculosis. An essential role for NAD in nonreplicating bacilli
J. Biol. Chem.
283
19329-19341
2008
Mycobacterium tuberculosis
Manually annotated by BRENDA team
LaRonde-LeBlanc, N.; Resto, M.; Gerratana, B.
Regulation of active site coupling in glutamine-dependent NAD(+) synthetase
Nat. Struct. Mol. Biol.
16
421-429
2009
Mycobacterium tuberculosis
Manually annotated by BRENDA team
Chuenchor, W.; Doukov, T.I.; Resto, M.; Chang, A.; Gerratana, B.
Regulation of the intersubunit ammonia tunnel in Mycobacterium tuberculosis glutamine-dependent NAD+ synthetase
Biochem. J.
443
417-426
2012
Mycobacterium tuberculosis (P9WJJ3), Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv (P9WJJ3)
Manually annotated by BRENDA team