BRENDA - Enzyme Database show

Biochemical studies and crystal structure determination of dihydrodipicolinate synthase from Pseudomonas aeruginosa

Kaur, N.; Gautam, A.; Kumar, S.; Singh, A.; Singh, N.; Sharma, S.; Sharma, R.; Tewari, R.; Singh, T.P.; Int. J. Biol. Macromol. 48, 779-787 (2011)

Data extracted from this reference:

Application
EC Number
Application
Commentary
Organism
4.3.3.7
drug development
the intracellular enzyme dihydrodipicolinate synthase a potential drug target because it is essential for the growth of bacteria while it is absent in humans
Pseudomonas aeruginosa
Cloned(Commentary)
EC Number
Commentary
Organism
4.3.3.7
gene dapA, DNA and amino acid sequence determination and analysis, expression of His-tagged DHDPS in Escherichia coli strain BL21(DE3)
Pseudomonas aeruginosa
Crystallization (Commentary)
EC Number
Crystallization
Organism
4.3.3.7
purified recombinant DHDPS, free or in complex with inhibitor (S)-lysine, 15 mg/ml protein in 50 mM Tris-HCl, pH 8.5, at 20C using hanging drop vapour diffusion method, mixing of 0.005 ml protein solution with 0.005 ml well solution containing 30% w/v PEG-3350, 170 mM MgCl2, 70 mM Tris-HCl, pH 8.5, and 6% v/v propylene glycol. Crystals obtained without 6% propylene glycol are soaked in the reservoir containing 20 mg/ml (S)-lysine, X-ray diffraction structure determination and analysis at 2.65-2.85 A resolution
Pseudomonas aeruginosa
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
4.3.3.7
L-lysine
binds at three sites to the enzyme, binding structure, overview
Pseudomonas aeruginosa
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.3.3.7
0.17
-
L-aspartate 4-semialdehyde
pH 8.0, 37C, recombinant His-tagged enzyme
Pseudomonas aeruginosa
4.3.3.7
0.9
-
pyruvate
pH 8.0, 37C, recombinant His-tagged enzyme
Pseudomonas aeruginosa
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
4.3.3.7
intracellular
-
Pseudomonas aeruginosa
5622
-
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
4.3.3.7
L-aspartate 4-semialdehyde + pyruvate
Pseudomonas aeruginosa
-
(S)-2,3-dihydropyridine-2,6-dicarboxylate + 2 H2O
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
4.3.3.7
no activity in Homo sapiens
-
-
-
4.3.3.7
Pseudomonas aeruginosa
Q9I4W3
gene dapA
-
Purification (Commentary)
EC Number
Commentary
Organism
4.3.3.7
recombinant His-tagged DHDPS from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
Pseudomonas aeruginosa
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
-
715195
Pseudomonas aeruginosa
(S)-2,3-dihydropyridine-2,6-dicarboxylate + 2 H2O
-
-
-
?
4.3.3.7
additional information
the active site is formed by residues Thr44, Tyr107 and Tyr133, stereochemically suitable for catalytic function
715195
Pseudomonas aeruginosa
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
Organism
4.3.3.7
dimer
three-dimensional structure determination shows that DHDPS forms a homodimer which is stabilized by several hydrogen bonds and van der Waals forces at the interface, active site structure, overview. Each monomer is composed of two domains, the N-terminal domain consists of residues 1-224, and forms an 8-fold parallel alpha/beta barrel
Pseudomonas aeruginosa
4.3.3.7
More
the recombinant enzyme adopts a characteristic alpha/beta conformation which is retained up to 65C
Pseudomonas aeruginosa
Temperature Optimum [C]
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
4.3.3.7
37
-
-
Pseudomonas aeruginosa
Temperature Stability [C]
EC Number
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
4.3.3.7
65
-
the recombinant enzyme adopts a characteristic alpha/beta conformation which is retained up to 65C
Pseudomonas aeruginosa
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
4.3.3.7
8
-
-
Pseudomonas aeruginosa
Application (protein specific)
EC Number
Application
Commentary
Organism
4.3.3.7
drug development
the intracellular enzyme dihydrodipicolinate synthase a potential drug target because it is essential for the growth of bacteria while it is absent in humans
Pseudomonas aeruginosa
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
4.3.3.7
gene dapA, DNA and amino acid sequence determination and analysis, expression of His-tagged DHDPS in Escherichia coli strain BL21(DE3)
Pseudomonas aeruginosa
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
4.3.3.7
purified recombinant DHDPS, free or in complex with inhibitor (S)-lysine, 15 mg/ml protein in 50 mM Tris-HCl, pH 8.5, at 20C using hanging drop vapour diffusion method, mixing of 0.005 ml protein solution with 0.005 ml well solution containing 30% w/v PEG-3350, 170 mM MgCl2, 70 mM Tris-HCl, pH 8.5, and 6% v/v propylene glycol. Crystals obtained without 6% propylene glycol are soaked in the reservoir containing 20 mg/ml (S)-lysine, X-ray diffraction structure determination and analysis at 2.65-2.85 A resolution
Pseudomonas aeruginosa
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
4.3.3.7
L-lysine
binds at three sites to the enzyme, binding structure, overview
Pseudomonas aeruginosa
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.3.3.7
0.17
-
L-aspartate 4-semialdehyde
pH 8.0, 37C, recombinant His-tagged enzyme
Pseudomonas aeruginosa
4.3.3.7
0.9
-
pyruvate
pH 8.0, 37C, recombinant His-tagged enzyme
Pseudomonas aeruginosa
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
4.3.3.7
intracellular
-
Pseudomonas aeruginosa
5622
-
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
4.3.3.7
L-aspartate 4-semialdehyde + pyruvate
Pseudomonas aeruginosa
-
(S)-2,3-dihydropyridine-2,6-dicarboxylate + 2 H2O
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
4.3.3.7
recombinant His-tagged DHDPS from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
Pseudomonas aeruginosa
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
-
715195
Pseudomonas aeruginosa
(S)-2,3-dihydropyridine-2,6-dicarboxylate + 2 H2O
-
-
-
?
4.3.3.7
additional information
the active site is formed by residues Thr44, Tyr107 and Tyr133, stereochemically suitable for catalytic function
715195
Pseudomonas aeruginosa
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
4.3.3.7
dimer
three-dimensional structure determination shows that DHDPS forms a homodimer which is stabilized by several hydrogen bonds and van der Waals forces at the interface, active site structure, overview. Each monomer is composed of two domains, the N-terminal domain consists of residues 1-224, and forms an 8-fold parallel alpha/beta barrel
Pseudomonas aeruginosa
4.3.3.7
More
the recombinant enzyme adopts a characteristic alpha/beta conformation which is retained up to 65C
Pseudomonas aeruginosa
Temperature Optimum [C] (protein specific)
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
4.3.3.7
37
-
-
Pseudomonas aeruginosa
Temperature Stability [C] (protein specific)
EC Number
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
4.3.3.7
65
-
the recombinant enzyme adopts a characteristic alpha/beta conformation which is retained up to 65C
Pseudomonas aeruginosa
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
4.3.3.7
8
-
-
Pseudomonas aeruginosa