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Literature summary for 4.3.3.7 extracted from

  • Dobson, R.C.; Valegard, K.; Gerrard, J.A.
    The crystal structure of three site-directed mutants of Escherichia coli dihydrodipicolinate synthase: further evidence for a catalytic triad (2004), J. Mol. Biol., 338, 329-339.
    View publication on PubMed
Search for more literature for 4.3.3.7

Cloned(Commentary)

Cloned (Comment) Organism
overexpression of wild-type and mutant enzymes in strain XL1-Blue Escherichia coli

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant wild-type and mutant enzymes, hanging or sitting drop vapour diffusion method, at 4°C and 21°C, 0.006 ml protein solutions: about 10 mg/ml protein, 1.8 M K2PO4, pH 10.0, N-octyl-beta-R-glucopyranoside 6% w/v, + 2 ml reservoir solution: 1.8 M K2PO4, pH 10.0, 3-4 days, X-ray diffraction structure determination and analysis at 2.35-2.5 A resolution Escherichia coli

Protein Variants

Protein Variants Comment Organism
additional information detailed structural properties of the mutant enzymes based on the crystal structure models Escherichia coli
T44V site-directed mutagenesis, reduced activity, structure is similar to the wild-type enzyme Escherichia coli
Y107F site-directed mutagenesis, reduced activity, structure is similar to the wild-type enzyme Escherichia coli
Y133F site-directed mutagenesis, reduced activity, structure is similar to the wild-type enzyme Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.08
-
 pyruvate mutant T44V, pH 8.0, 30°C Escherichia coli
0.09
-
 (S)-aspartate 4-semialdehyde mutant T44V, pH 8.0, 30°C Escherichia coli
0.11
-
 (S)-aspartate 4-semialdehyde wild-type enzyme, pH 8.0, 30°C Escherichia coli
0.16
-
 pyruvate mutant Y107F, pH 8.0, 30°C Escherichia coli
0.26
-
 pyruvate wild-type enzyme, pH 8.0, 30°C Escherichia coli
0.58
-
 (S)-aspartate 4-semialdehyde mutant Y107F, pH 8.0, 30°C Escherichia coli
2.7
-
 (S)-aspartate 4-semialdehyde mutant Y133F, pH 8.0, 30°C Escherichia coli
35
-
 pyruvate mutant Y133F, pH 8.0, 30°C Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(S)-aspartate 4-semialdehyde + pyruvate Escherichia coli branch point reaction in the biosynthesis of lysine dihydrodipicolinate + H2O
-
 ?

Organism

Organism UniProt Comment Textmining
Escherichia coli P0A6L2
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type 5.8fold, recombinant mutant Y107F 17.6fold, recombinant mutant T44V 15.8fold, and recombinant mutant Y133F 18fold Escherichia coli

Reaction

Reaction Comment Organism Reaction ID
pyruvate + L-aspartate-4-semialdehyde = (2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinate + H2O ping-pong kinetic and catalytic mechanism involves a catalytic triad which consists of Tyr133, Thr44, and Tyr107, overview Escherichia coli
a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.0034
-
 purified mutant T44V Escherichia coli
0.008
-
 purified mutant Y133F Escherichia coli
0.27
-
 purified mutant Y107F Escherichia coli
1.81
-
 purified wild-type enzyme Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(S)-aspartate 4-semialdehyde + pyruvate
-
 Escherichia coli dihydrodipicolinate + H2O
-
 ?
(S)-aspartate 4-semialdehyde + pyruvate branch point reaction in the biosynthesis of lysine Escherichia coli dihydrodipicolinate + H2O
-
 ?

Subunits

Subunits Comment Organism
tetramer homotetramer of (alpha/beta)8 barrels, each containing one active site, crystal structure Escherichia coli

Synonyms

Synonyms Comment Organism
DHDPS
-
 Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.12
-
 (S)-aspartate 4-semialdehyde mutant T44V, pH 8.0, 30°C Escherichia coli
0.7
-
 (S)-aspartate 4-semialdehyde mutant Y133F, pH 8.0, 30°C Escherichia coli
10.8
-
 (S)-aspartate 4-semialdehyde mutant Y107F, pH 8.0, 30°C Escherichia coli
124
-
 (S)-aspartate 4-semialdehyde wild-type enzyme, pH 8.0, 30°C Escherichia coli