BRENDA - Enzyme Database show

Exploring the dihydrodipicolinate synthase tetramer: how resilient is the dimer-dimer interface?

Griffin, M.D.; Dobson, R.C.; Gerrard, J.A.; Perugini, M.A.; Arch. Biochem. Biophys. 494, 58-63 (2010)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
4.3.3.7
DHDPS mutants expressed in Escherichia coli strain AT997r-
Escherichia coli
Engineering
EC Number
Amino acid exchange
Commentary
Organism
4.3.3.7
D193A
removal of water mediated hydrogen-bond network
Escherichia coli
4.3.3.7
D193Y
removal of water mediated hydrogen-bond network and introduction of steric bulk to alter surface topology
Escherichia coli
4.3.3.7
Q196D
removal of hydrogen bonds and charge-charge repulsion, shortened side chain places charged carboxyl groups proximal at interface
Escherichia coli
4.3.3.7
Q234D
removal of hydrogen bond and charge-charge repulsion with negatively charged E175. Quaternary structure appears closest to that of the wild-type enzyme
Escherichia coli
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
4.3.3.7
L-lysine
-
Escherichia coli
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.3.3.7
0.13
-
L-aspartate 4-semialdehyde
wild-type, at 30C
Escherichia coli
4.3.3.7
0.15
-
L-aspartate 4-semialdehyde
mutants Q196D, D193A and D193Y, at 30C
Escherichia coli
4.3.3.7
0.16
-
pyruvate
wild-type, at 30C
Escherichia coli
4.3.3.7
0.17
-
L-aspartate 4-semialdehyde
mutant D193A, at 30C
Escherichia coli
4.3.3.7
0.32
-
pyruvate
mutant Q196D, at 30C
Escherichia coli
4.3.3.7
0.44
-
pyruvate
mutant D193A, at 30C
Escherichia coli
4.3.3.7
0.46
-
pyruvate
mutant Q234D, at 30C
Escherichia coli
4.3.3.7
0.57
-
pyruvate
mutant D193Y, at 30C
Escherichia coli
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
4.3.3.7
Escherichia coli
P0A6L2
-
-
Purification (Commentary)
EC Number
Commentary
Organism
4.3.3.7
wild-type DHDPS, and the coupling enzyme, DHDPR, by ammonium sulphate fractionation
Escherichia coli
Storage Stability
EC Number
Storage Stability
Organism
4.3.3.7
-20C, 20 mM Tris-HCl buffer, pH 8.0
Escherichia coli
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
-
701934
Escherichia coli
dihydrodipicolinate + 2 H2O
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
4.3.3.7
tetramer
the dimer-dimer interface is small, accounts only 4.3% of the subunit surface area
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
6.5
-
L-aspartate 4-semialdehyde
mutant D193Y, at 30C
Escherichia coli
4.3.3.7
7.2
-
pyruvate
mutant D193Y, at 30C
Escherichia coli
4.3.3.7
9.15
-
pyruvate
mutant D193A, at 30C
Escherichia coli
4.3.3.7
11.8
-
pyruvate
mutant Q234D, at 30C
Escherichia coli
4.3.3.7
12.4
-
L-aspartate 4-semialdehyde
mutant D193A, at 30C
Escherichia coli
4.3.3.7
13.28
-
pyruvate
mutant Q196D, at 30C
Escherichia coli
4.3.3.7
78
-
pyruvate
wild-type, at 30C
Escherichia coli
Ki Value [mM]
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
4.3.3.7
0.18
-
L-lysine
mutant D193A, with respect to (S)-aspartate 4-semialdehyde
Escherichia coli
4.3.3.7
0.19
-
L-lysine
mutant Q196D, with respect to (S)-aspartate 4-semialdehyde
Escherichia coli
4.3.3.7
0.4
-
L-lysine
wild-type, with respect to (S)-aspartate 4-semialdehyde
Escherichia coli
4.3.3.7
2.2
-
L-lysine
mutant D193A, with respect to pyruvate
Escherichia coli
4.3.3.7
3.6
-
L-lysine
wild-type, with respect to pyruvate
Escherichia coli
4.3.3.7
4.1
-
L-lysine
mutant Q196D, with respect to pyruvate
Escherichia coli
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
4.3.3.7
DHDPS mutants expressed in Escherichia coli strain AT997r-
Escherichia coli
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
4.3.3.7
D193A
removal of water mediated hydrogen-bond network
Escherichia coli
4.3.3.7
D193Y
removal of water mediated hydrogen-bond network and introduction of steric bulk to alter surface topology
Escherichia coli
4.3.3.7
Q196D
removal of hydrogen bonds and charge-charge repulsion, shortened side chain places charged carboxyl groups proximal at interface
Escherichia coli
4.3.3.7
Q234D
removal of hydrogen bond and charge-charge repulsion with negatively charged E175. Quaternary structure appears closest to that of the wild-type enzyme
Escherichia coli
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
4.3.3.7
L-lysine
-
Escherichia coli
Ki Value [mM] (protein specific)
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
4.3.3.7
0.18
-
L-lysine
mutant D193A, with respect to (S)-aspartate 4-semialdehyde
Escherichia coli
4.3.3.7
0.19
-
L-lysine
mutant Q196D, with respect to (S)-aspartate 4-semialdehyde
Escherichia coli
4.3.3.7
0.4
-
L-lysine
wild-type, with respect to (S)-aspartate 4-semialdehyde
Escherichia coli
4.3.3.7
2.2
-
L-lysine
mutant D193A, with respect to pyruvate
Escherichia coli
4.3.3.7
3.6
-
L-lysine
wild-type, with respect to pyruvate
Escherichia coli
4.3.3.7
4.1
-
L-lysine
mutant Q196D, with respect to pyruvate
Escherichia coli
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.3.3.7
0.13
-
L-aspartate 4-semialdehyde
wild-type, at 30C
Escherichia coli
4.3.3.7
0.15
-
L-aspartate 4-semialdehyde
mutants Q196D, D193A and D193Y, at 30C
Escherichia coli
4.3.3.7
0.16
-
pyruvate
wild-type, at 30C
Escherichia coli
4.3.3.7
0.17
-
L-aspartate 4-semialdehyde
mutant D193A, at 30C
Escherichia coli
4.3.3.7
0.32
-
pyruvate
mutant Q196D, at 30C
Escherichia coli
4.3.3.7
0.44
-
pyruvate
mutant D193A, at 30C
Escherichia coli
4.3.3.7
0.46
-
pyruvate
mutant Q234D, at 30C
Escherichia coli
4.3.3.7
0.57
-
pyruvate
mutant D193Y, at 30C
Escherichia coli
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
4.3.3.7
wild-type DHDPS, and the coupling enzyme, DHDPR, by ammonium sulphate fractionation
Escherichia coli
Storage Stability (protein specific)
EC Number
Storage Stability
Organism
4.3.3.7
-20C, 20 mM Tris-HCl buffer, pH 8.0
Escherichia coli
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
-
701934
Escherichia coli
dihydrodipicolinate + 2 H2O
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
4.3.3.7
tetramer
the dimer-dimer interface is small, accounts only 4.3% of the subunit surface area
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
6.5
-
L-aspartate 4-semialdehyde
mutant D193Y, at 30C
Escherichia coli
4.3.3.7
7.2
-
pyruvate
mutant D193Y, at 30C
Escherichia coli
4.3.3.7
9.15
-
pyruvate
mutant D193A, at 30C
Escherichia coli
4.3.3.7
11.8
-
pyruvate
mutant Q234D, at 30C
Escherichia coli
4.3.3.7
12.4
-
L-aspartate 4-semialdehyde
mutant D193A, at 30C
Escherichia coli
4.3.3.7
13.28
-
pyruvate
mutant Q196D, at 30C
Escherichia coli
4.3.3.7
78
-
pyruvate
wild-type, at 30C
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
12.6
-
pyruvate
mutant D193Y, at 30C
Escherichia coli
4.3.3.7
21
-
pyruvate
mutant D193A, at 30C
Escherichia coli
4.3.3.7
25.6
-
pyruvate
mutant Q234D, at 30C
Escherichia coli
4.3.3.7
42
-
pyruvate
mutant Q196D, at 30C
Escherichia coli
4.3.3.7
43
-
L-aspartate 4-semialdehyde
mutant D193Y, at 30C
Escherichia coli
4.3.3.7
53
-
L-aspartate 4-semialdehyde
mutant D193A, at 30C
Escherichia coli
4.3.3.7
83
-
L-aspartate 4-semialdehyde
mutant D193A, at 30C
Escherichia coli
4.3.3.7
88
-
L-aspartate 4-semialdehyde
mutant Q196D, at 30C
Escherichia coli
4.3.3.7
488
-
pyruvate
wild-type, at 30C
Escherichia coli
4.3.3.7
600
-
L-aspartate 4-semialdehyde
wild-type, at 30C
Escherichia coli
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
12.6
-
pyruvate
mutant D193Y, at 30C
Escherichia coli
4.3.3.7
21
-
pyruvate
mutant D193A, at 30C
Escherichia coli
4.3.3.7
25.6
-
pyruvate
mutant Q234D, at 30C
Escherichia coli
4.3.3.7
42
-
pyruvate
mutant Q196D, at 30C
Escherichia coli
4.3.3.7
43
-
L-aspartate 4-semialdehyde
mutant D193Y, at 30C
Escherichia coli
4.3.3.7
53
-
L-aspartate 4-semialdehyde
mutant D193A, at 30C
Escherichia coli
4.3.3.7
83
-
L-aspartate 4-semialdehyde
mutant D193A, at 30C
Escherichia coli
4.3.3.7
88
-
L-aspartate 4-semialdehyde
mutant Q196D, at 30C
Escherichia coli
4.3.3.7
488
-
pyruvate
wild-type, at 30C
Escherichia coli
4.3.3.7
600
-
L-aspartate 4-semialdehyde
wild-type, at 30C
Escherichia coli