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Literature summary for 4.3.3.7 extracted from

  • 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.
    A tetrameric structure is not essential for activity in dihydrodipicolinate synthase (DHDPS) from Mycobacterium tuberculosis (2011), Arch. Biochem. Biophys., 512, 154-159.
    View publication on PubMed

Crystallization (Commentary)

Crystallization (Comment) Organism
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

Protein Variants

Protein Variants Comment Organism
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]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.17
-
pyruvate pH 8.0, 30°C, recombinant wild-type enzyme Mycobacterium tuberculosis
0.33
-
pyruvate pH 8.0, 30°C, recombinant mutant A204R Mycobacterium tuberculosis
0.43
-
L-aspartate 4-semialdehyde pH 8.0, 30°C, recombinant wild-type enzyme Mycobacterium tuberculosis
1.1
-
L-aspartate 4-semialdehyde pH 8.0, 30°C, recombinant mutant A204R Mycobacterium tuberculosis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
62400
-
2 * 62400, mutant A204R, crystal structure Mycobacterium tuberculosis
62400
-
4 * 62400, crystal structure, the tetrameric structure is not essential for activity in DHDPS from Mycobacterium tuberculosis Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-aspartate 4-semialdehyde + pyruvate Mycobacterium tuberculosis
-
(S)-2,3-dihydropyridine-2,6-dicarboxylate + 2 H2O
-
?

Organism

Organism UniProt Comment Textmining
Mycobacterium tuberculosis
-
-
-

Reaction

Reaction Comment Organism Reaction ID
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)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-aspartate 4-semialdehyde + pyruvate
-
Mycobacterium tuberculosis (S)-2,3-dihydropyridine-2,6-dicarboxylate + 2 H2O
-
?

Subunits

Subunits Comment Organism
dimer 2 * 62400, mutant A204R, crystal structure Mycobacterium tuberculosis
tetramer 4 * 62400, crystal structure, the tetrameric structure is not essential for activity in DHDPS from Mycobacterium tuberculosis Mycobacterium tuberculosis

Synonyms

Synonyms Comment Organism
DHDPS
-
Mycobacterium tuberculosis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Mycobacterium tuberculosis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
119
-
pyruvate pH 8.0, 30°C, recombinant mutant A204R Mycobacterium tuberculosis
119
-
L-aspartate 4-semialdehyde pH 8.0, 30°C, recombinant mutant A204R Mycobacterium tuberculosis
138
-
pyruvate pH 8.0, 30°C, recombinant wild-type enzyme Mycobacterium tuberculosis
138
-
L-aspartate 4-semialdehyde pH 8.0, 30°C, recombinant wild-type enzyme Mycobacterium tuberculosis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.75 8.25
-
Mycobacterium tuberculosis

General Information

General Information Comment Organism
additional information the tetrameric structure is not essential for activity in DHDPS from Mycobacterium tuberculosis Mycobacterium tuberculosis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
110
-
L-aspartate 4-semialdehyde pH 8.0, 30°C, recombinant mutant A204R Mycobacterium tuberculosis
320
-
L-aspartate 4-semialdehyde pH 8.0, 30°C, recombinant wild-type enzyme Mycobacterium tuberculosis
360
-
pyruvate pH 8.0, 30°C, recombinant mutant A204R Mycobacterium tuberculosis
820
-
pyruvate pH 8.0, 30°C, recombinant wild-type enzyme Mycobacterium tuberculosis