BRENDA - Enzyme Database show

Cloning and characterisation of dihydrodipicolinate synthase from the pathogen Neisseria meningitidis

Devenish, S.R.; Huisman, F.H.; Parker, E.J.; Hadfield, A.T.; Gerrard, J.A.; Biochim. Biophys. Acta 1794, 1168-1174 (2009)

Data extracted from this reference:

Application
EC Number
Application
Commentary
Organism
4.3.3.7
drug development
the allosteric binding site of DHDPS may be a good starting point for development of an inhibitor specific to Neisseria meningitidis
Neisseria meningitidis
Cloned(Commentary)
EC Number
Commentary
Organism
4.3.3.7
as His-tagged constructs
Escherichia coli
4.3.3.7
into the expression vector pET151-D to give the plasmid pFH02, to transform chemically super-competent Escherichia coli Top10 cells
Neisseria meningitidis
Crystallization (Commentary)
EC Number
Crystallization
Organism
4.3.3.7
to 2.0 A resolution, space group of the crystal is P212121, with unit cell dimensions of a = 80.7, b = 115.7 and c = 132.1 A. The secondary and tertiary structures are remarkably similar to that of Escherichia coli DHDPS. The hydrogen bond lengths within the catalytic triad, and particularly that between Y133 and T44, differ significantly from those of the Escherichia coli enzyme
Neisseria meningitidis
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
4.3.3.7
L-aspartate 4-semialdehyde
is subject to substrate inhibition by high concentrations
Neisseria meningitidis
4.3.3.7
L-lysine
-
Escherichia coli
4.3.3.7
L-lysine
is significantly more sensitive to feedback inhibition than Escherichia coli DHDPS
Neisseria meningitidis
4.3.3.7
additional information
no substrate inhibition by (S)-aspartate 4-semialdehyde
Escherichia coli
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.3.3.7
0.052
-
L-aspartate 4-semialdehyde
at 30°C, in 100 mM HEPES buffer, pH 8.0
Neisseria meningitidis
4.3.3.7
0.11
-
L-aspartate 4-semialdehyde
at 30°C, in 100 mM HEPES buffer, pH 8.0
Escherichia coli
4.3.3.7
0.26
-
pyruvate
at 30°C, in 100 mM HEPES buffer, pH 8.0
Escherichia coli
4.3.3.7
0.5
-
pyruvate
at 30°C, in 100 mM HEPES buffer, pH 8.0
Neisseria meningitidis
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
4.3.3.7
31510
-
mass spectrometry
Neisseria meningitidis
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
4.3.3.7
Escherichia coli
-
-
-
4.3.3.7
Neisseria meningitidis
Q9JZR4
-
-
4.3.3.7
Neisseria meningitidis MC58
Q9JZR4
-
-
Purification (Commentary)
EC Number
Commentary
Organism
4.3.3.7
by centrifugation and gel filtration
Escherichia coli
4.3.3.7
by centrifugation and gel filtration, to homogeneity
Neisseria meningitidis
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4.3.3.7
L-aspartate 4-semialdehyde + pyruvate
-
702445
Escherichia coli
dihydrodipicolinate + 2 H2O
-
-
-
?
4.3.3.7
L-aspartate 4-semialdehyde + pyruvate
slightly higher affinity for L-aspartate 4-semialdehyde and somewhat lower affinity for pyruvate than the Escherichia coli enzyme, whereby the catalytic activity is similar
702445
Neisseria meningitidis
dihydrodipicolinate + 2 H2O
-
-
-
?
4.3.3.7
L-aspartate 4-semialdehyde + pyruvate
slightly higher affinity for L-aspartate 4-semialdehyde and somewhat lower affinity for pyruvate than the Escherichia coli enzyme, whereby the catalytic activity is similar
702445
Neisseria meningitidis MC58
dihydrodipicolinate + 2 H2O
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
4.3.3.7
homotetramer
crystallography
Neisseria meningitidis
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
4.3.3.7
46.5
-
melting temperature in the absence of substrates
Neisseria meningitidis
4.3.3.7
59.5
-
melting temperature in the absence of substrates
Escherichia coli
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
4.3.3.7
46.7
-
pyruvate
at 30°C, in 100 mM HEPES buffer, pH 8.0
Neisseria meningitidis
4.3.3.7
124
-
pyruvate
at 30°C, in 100 mM HEPES buffer, pH 8.0
Escherichia coli
Ki Value [mM]
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
4.3.3.7
0.058
-
L-lysine
with (S)-aspartate 4-semialdehyde as substrate
Neisseria meningitidis
4.3.3.7
0.32
-
L-lysine
with (S)-aspartate 4-semialdehyde as substrate
Escherichia coli
4.3.3.7
1.7
-
L-lysine
with pyruvate as substrate
Neisseria meningitidis
4.3.3.7
3.9
-
L-lysine
with pyruvate as substrate
Escherichia coli
IC50 Value
EC Number
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
4.3.3.7
0.053
-
-
Neisseria meningitidis
L-lysine
Application (protein specific)
EC Number
Application
Commentary
Organism
4.3.3.7
drug development
the allosteric binding site of DHDPS may be a good starting point for development of an inhibitor specific to Neisseria meningitidis
Neisseria meningitidis
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
4.3.3.7
as His-tagged constructs
Escherichia coli
4.3.3.7
into the expression vector pET151-D to give the plasmid pFH02, to transform chemically super-competent Escherichia coli Top10 cells
Neisseria meningitidis
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
4.3.3.7
to 2.0 A resolution, space group of the crystal is P212121, with unit cell dimensions of a = 80.7, b = 115.7 and c = 132.1 A. The secondary and tertiary structures are remarkably similar to that of Escherichia coli DHDPS. The hydrogen bond lengths within the catalytic triad, and particularly that between Y133 and T44, differ significantly from those of the Escherichia coli enzyme
Neisseria meningitidis
IC50 Value (protein specific)
EC Number
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
4.3.3.7
0.053
-
-
Neisseria meningitidis
L-lysine
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
4.3.3.7
L-aspartate 4-semialdehyde
is subject to substrate inhibition by high concentrations
Neisseria meningitidis
4.3.3.7
L-lysine
-
Escherichia coli
4.3.3.7
L-lysine
is significantly more sensitive to feedback inhibition than Escherichia coli DHDPS
Neisseria meningitidis
4.3.3.7
additional information
no substrate inhibition by (S)-aspartate 4-semialdehyde
Escherichia coli
Ki Value [mM] (protein specific)
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
4.3.3.7
0.058
-
L-lysine
with (S)-aspartate 4-semialdehyde as substrate
Neisseria meningitidis
4.3.3.7
0.32
-
L-lysine
with (S)-aspartate 4-semialdehyde as substrate
Escherichia coli
4.3.3.7
1.7
-
L-lysine
with pyruvate as substrate
Neisseria meningitidis
4.3.3.7
3.9
-
L-lysine
with pyruvate as substrate
Escherichia coli
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.3.3.7
0.052
-
L-aspartate 4-semialdehyde
at 30°C, in 100 mM HEPES buffer, pH 8.0
Neisseria meningitidis
4.3.3.7
0.11
-
L-aspartate 4-semialdehyde
at 30°C, in 100 mM HEPES buffer, pH 8.0
Escherichia coli
4.3.3.7
0.26
-
pyruvate
at 30°C, in 100 mM HEPES buffer, pH 8.0
Escherichia coli
4.3.3.7
0.5
-
pyruvate
at 30°C, in 100 mM HEPES buffer, pH 8.0
Neisseria meningitidis
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
4.3.3.7
31510
-
mass spectrometry
Neisseria meningitidis
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
4.3.3.7
by centrifugation and gel filtration
Escherichia coli
4.3.3.7
by centrifugation and gel filtration, to homogeneity
Neisseria meningitidis
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4.3.3.7
L-aspartate 4-semialdehyde + pyruvate
-
702445
Escherichia coli
dihydrodipicolinate + 2 H2O
-
-
-
?
4.3.3.7
L-aspartate 4-semialdehyde + pyruvate
slightly higher affinity for L-aspartate 4-semialdehyde and somewhat lower affinity for pyruvate than the Escherichia coli enzyme, whereby the catalytic activity is similar
702445
Neisseria meningitidis
dihydrodipicolinate + 2 H2O
-
-
-
?
4.3.3.7
L-aspartate 4-semialdehyde + pyruvate
slightly higher affinity for L-aspartate 4-semialdehyde and somewhat lower affinity for pyruvate than the Escherichia coli enzyme, whereby the catalytic activity is similar
702445
Neisseria meningitidis MC58
dihydrodipicolinate + 2 H2O
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
4.3.3.7
homotetramer
crystallography
Neisseria meningitidis
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
4.3.3.7
46.5
-
melting temperature in the absence of substrates
Neisseria meningitidis
4.3.3.7
59.5
-
melting temperature in the absence of substrates
Escherichia coli
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
4.3.3.7
46.7
-
pyruvate
at 30°C, in 100 mM HEPES buffer, pH 8.0
Neisseria meningitidis
4.3.3.7
124
-
pyruvate
at 30°C, in 100 mM HEPES buffer, pH 8.0
Escherichia coli
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
4.3.3.7
93.4
-
pyruvate
at 30°C, in 100 mM HEPES buffer, pH 8.0
Neisseria meningitidis
4.3.3.7
898
-
L-aspartate 4-semialdehyde
at 30°C, in 100 mM HEPES buffer, pH 8.0
Neisseria meningitidis
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
4.3.3.7
93.4
-
pyruvate
at 30°C, in 100 mM HEPES buffer, pH 8.0
Neisseria meningitidis
4.3.3.7
898
-
L-aspartate 4-semialdehyde
at 30°C, in 100 mM HEPES buffer, pH 8.0
Neisseria meningitidis