BRENDA - Enzyme Database show

A tetrameric structure is not essential for activity in dihydrodipicolinate synthase (DHDPS) from Mycobacterium tuberculosis

Evans, G.; Schuldt, L.; Griffin, M.D.; Devenish, S.R.; Grant Pearce, F.; Perugini, M.A.; Dobson, R.C.; Jameson, G.B.; Weiss, M.S.; Gerrard, J.A.; Arch. Biochem. Biophys. 512, 154-159 (2011)

Data extracted from this reference:

Crystallization (Commentary)
EC Number
Crystallization
Organism
4.3.3.7
purified recombinant DHDPS mutant A204R, sitting drop vapour diffusion method at room temperature, 200 nl of 10 mg/ml protein in 20 mM Tris-HCl, 2 mM 2-mercaptoethanol, 250 mM NaCl, 5% v/v glycerol, 10 mM pyruvate, pH 8.0, is mixed with 230 nl reservoir solution containing 2.0 M ammonium sulphate, 100 mM sodium acetate, pH 5.5, X-ray diffraction structure determination and analysis at 2.0 A resolution
Mycobacterium tuberculosis
Engineering
EC Number
Amino acid exchange
Commentary
Organism
4.3.3.7
A204R
site-directed mutagenesis, disruption of the native tetramer formation by targeting the dimer-dimer interface. The mutant enzyme is a dimeric protein with an identical fold and active-site structure to the tetrameric wild-type enzyme
Mycobacterium tuberculosis
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.3.3.7
0.17
-
pyruvate
pH 8.0, 30°C, recombinant wild-type enzyme
Mycobacterium tuberculosis
4.3.3.7
0.33
-
pyruvate
pH 8.0, 30°C, recombinant mutant A204R
Mycobacterium tuberculosis
4.3.3.7
0.43
-
L-aspartate 4-semialdehyde
pH 8.0, 30°C, recombinant wild-type enzyme
Mycobacterium tuberculosis
4.3.3.7
1.1
-
L-aspartate 4-semialdehyde
pH 8.0, 30°C, recombinant mutant A204R
Mycobacterium tuberculosis
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
4.3.3.7
62400
-
2 * 62400, mutant A204R, crystal structure; 4 * 62400, crystal structure, the tetrameric structure is not essential for activity in DHDPS from Mycobacterium tuberculosis
Mycobacterium tuberculosis
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
Mycobacterium tuberculosis
-
(S)-2,3-dihydropyridine-2,6-dicarboxylate + 2 H2O
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
4.3.3.7
Mycobacterium tuberculosis
-
-
-
Reaction
EC Number
Reaction
Commentary
Organism
4.3.3.7
pyruvate + L-aspartate-4-semialdehyde = (2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinate + H2O
reaction mechanism including Schiff base formation, tautomerization, and cyclization, overview
Mycobacterium tuberculosis
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
-
713969
Mycobacterium tuberculosis
(S)-2,3-dihydropyridine-2,6-dicarboxylate + 2 H2O
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
4.3.3.7
dimer
2 * 62400, mutant A204R, crystal structure
Mycobacterium tuberculosis
4.3.3.7
tetramer
4 * 62400, crystal structure, the tetrameric structure is not essential for activity in DHDPS from Mycobacterium tuberculosis
Mycobacterium tuberculosis
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
4.3.3.7
30
-
assay at
Mycobacterium tuberculosis
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
4.3.3.7
119
-
L-aspartate 4-semialdehyde
pH 8.0, 30°C, recombinant mutant A204R
Mycobacterium tuberculosis
4.3.3.7
119
-
pyruvate
pH 8.0, 30°C, recombinant mutant A204R
Mycobacterium tuberculosis
4.3.3.7
138
-
L-aspartate 4-semialdehyde
pH 8.0, 30°C, recombinant wild-type enzyme
Mycobacterium tuberculosis
4.3.3.7
138
-
pyruvate
pH 8.0, 30°C, recombinant wild-type enzyme
Mycobacterium tuberculosis
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
4.3.3.7
7.75
8.25
-
Mycobacterium tuberculosis
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
4.3.3.7
purified recombinant DHDPS mutant A204R, sitting drop vapour diffusion method at room temperature, 200 nl of 10 mg/ml protein in 20 mM Tris-HCl, 2 mM 2-mercaptoethanol, 250 mM NaCl, 5% v/v glycerol, 10 mM pyruvate, pH 8.0, is mixed with 230 nl reservoir solution containing 2.0 M ammonium sulphate, 100 mM sodium acetate, pH 5.5, X-ray diffraction structure determination and analysis at 2.0 A resolution
Mycobacterium tuberculosis
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
4.3.3.7
A204R
site-directed mutagenesis, disruption of the native tetramer formation by targeting the dimer-dimer interface. The mutant enzyme is a dimeric protein with an identical fold and active-site structure to the tetrameric wild-type enzyme
Mycobacterium tuberculosis
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.3.3.7
0.17
-
pyruvate
pH 8.0, 30°C, recombinant wild-type enzyme
Mycobacterium tuberculosis
4.3.3.7
0.33
-
pyruvate
pH 8.0, 30°C, recombinant mutant A204R
Mycobacterium tuberculosis
4.3.3.7
0.43
-
L-aspartate 4-semialdehyde
pH 8.0, 30°C, recombinant wild-type enzyme
Mycobacterium tuberculosis
4.3.3.7
1.1
-
L-aspartate 4-semialdehyde
pH 8.0, 30°C, recombinant mutant A204R
Mycobacterium tuberculosis
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
4.3.3.7
62400
-
2 * 62400, mutant A204R, crystal structure; 4 * 62400, crystal structure, the tetrameric structure is not essential for activity in DHDPS from Mycobacterium tuberculosis
Mycobacterium tuberculosis
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
Mycobacterium tuberculosis
-
(S)-2,3-dihydropyridine-2,6-dicarboxylate + 2 H2O
-
-
?
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
-
713969
Mycobacterium tuberculosis
(S)-2,3-dihydropyridine-2,6-dicarboxylate + 2 H2O
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
4.3.3.7
dimer
2 * 62400, mutant A204R, crystal structure
Mycobacterium tuberculosis
4.3.3.7
tetramer
4 * 62400, crystal structure, the tetrameric structure is not essential for activity in DHDPS from Mycobacterium tuberculosis
Mycobacterium tuberculosis
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
4.3.3.7
30
-
assay at
Mycobacterium tuberculosis
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
119
-
L-aspartate 4-semialdehyde
pH 8.0, 30°C, recombinant mutant A204R
Mycobacterium tuberculosis
4.3.3.7
119
-
pyruvate
pH 8.0, 30°C, recombinant mutant A204R
Mycobacterium tuberculosis
4.3.3.7
138
-
L-aspartate 4-semialdehyde
pH 8.0, 30°C, recombinant wild-type enzyme
Mycobacterium tuberculosis
4.3.3.7
138
-
pyruvate
pH 8.0, 30°C, recombinant wild-type enzyme
Mycobacterium tuberculosis
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
4.3.3.7
7.75
8.25
-
Mycobacterium tuberculosis
General Information
EC Number
General Information
Commentary
Organism
4.3.3.7
additional information
the tetrameric structure is not essential for activity in DHDPS from Mycobacterium tuberculosis
Mycobacterium tuberculosis
General Information (protein specific)
EC Number
General Information
Commentary
Organism
4.3.3.7
additional information
the tetrameric structure is not essential for activity in DHDPS from Mycobacterium tuberculosis
Mycobacterium tuberculosis
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
110
-
L-aspartate 4-semialdehyde
pH 8.0, 30°C, recombinant mutant A204R
Mycobacterium tuberculosis
4.3.3.7
320
-
L-aspartate 4-semialdehyde
pH 8.0, 30°C, recombinant wild-type enzyme
Mycobacterium tuberculosis
4.3.3.7
360
-
pyruvate
pH 8.0, 30°C, recombinant mutant A204R
Mycobacterium tuberculosis
4.3.3.7
820
-
pyruvate
pH 8.0, 30°C, recombinant wild-type enzyme
Mycobacterium tuberculosis
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
110
-
L-aspartate 4-semialdehyde
pH 8.0, 30°C, recombinant mutant A204R
Mycobacterium tuberculosis
4.3.3.7
320
-
L-aspartate 4-semialdehyde
pH 8.0, 30°C, recombinant wild-type enzyme
Mycobacterium tuberculosis
4.3.3.7
360
-
pyruvate
pH 8.0, 30°C, recombinant mutant A204R
Mycobacterium tuberculosis
4.3.3.7
820
-
pyruvate
pH 8.0, 30°C, recombinant wild-type enzyme
Mycobacterium tuberculosis