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Literature summary for 4.3.3.7 extracted from

  • Pearce, F.G.; Dobson, R.C.; Jameson, G.B.; Perugini, M.A.; Gerrard, J.A.
    Characterization of monomeric dihydrodipicolinate synthase variant reveals the importance of substrate binding in optimizing oligomerization (2011), Biochim. Biophys. Acta, 1814, 1900-1909.
    View publication on PubMed
Search for more literature for 4.3.3.7

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant Tm-DHDPS-DELTAArg-237, vapor diffusion method, mixing of 150 nl protein solution, containing 11.2 mg/ml protein in 20 mM Tris-HCl, pH 8.0, with 150 nl reservoir solution, containing 40% v/v PEG 300, 100 mM phosphate-citrate, buffer, pH 4.2, and 0.02% w/v sodium azide, X-ray diffraction structure determination and analysis at 1.9-2.1 A resolution Thermotoga maritima

Protein Variants

Protein Variants Comment Organism
additional information construction of mutants Tm-DHDPS-DELTAAsp168, DELTAAsp171, or DELTAArg237 by mutating charged residues, reduction of the number of salt bridges at one of the two tetramerization interface of the enzyme and its interactions results in variants with altered quaternary structure, e.g. monomeric, as shown by analytical ultracentrifugation, gel filtration liquid chromatography, and small angle X-ray scattering, and X-ray crystallographic studies, overview Thermotoga maritima

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information Michaelis-Menten kinetics for wild-type and mutant enzymes, overview Thermotoga maritima
0.05
-
 pyruvate pH not specified in the publication, 30°C, recombinant mutant DELTAAsp237 Thermotoga maritima
0.07
-
 pyruvate pH not specified in the publication, 30°C, recombinant mutant DELTAAsp171 Thermotoga maritima
0.08
-
 pyruvate pH not specified in the publication, 30°C, recombinant mutant DELTAAsp171/Arg237 Thermotoga maritima
0.08
-
 pyruvate pH not specified in the publication, 30°C, recombinant wild-type enzyme Thermotoga maritima
0.1
-
 pyruvate pH not specified in the publication, 45°C, recombinant mutant DELTAAsp237 Thermotoga maritima
0.15
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 45°C, recombinant mutant DELTAAsp237 Thermotoga maritima
0.15
-
 pyruvate pH not specified in the publication, 45°C, recombinant wild-type enzyme Thermotoga maritima
0.16
-
 pyruvate pH not specified in the publication, 45°C, recombinant mutant DELTAAsp171 Thermotoga maritima
0.18
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 30°C, recombinant mutant DELTAAsp237 Thermotoga maritima
0.18
-
 pyruvate pH not specified in the publication, 45°C, recombinant mutant DELTAAsp171/Arg237 Thermotoga maritima
0.23
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 30°C, recombinant mutant DELTAAsp171/Arg237 Thermotoga maritima
0.23
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 30°C, recombinant wild-type enzyme Thermotoga maritima
0.32
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 45°C, recombinant mutant DELTAAsp171/Arg237 Thermotoga maritima
0.36
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 45°C, recombinant wild-type enzyme Thermotoga maritima
0.42
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 30°C, recombinant mutant DELTAAsp171 Thermotoga maritima
0.46
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 45°C, recombinant mutant DELTAAsp171 Thermotoga maritima
1.1
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 45°C, recombinant mutant DELTAAsp168/Asp171 Thermotoga maritima
1.2
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 30°C, recombinant mutant DELTAAsp168/Asp237 Thermotoga maritima
1.3
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 30°C, recombinant mutant DELTAAsp168/Asp171 Thermotoga maritima
1.5
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 30°C, recombinant mutant DELTAAsp168 Thermotoga maritima
1.5
-
 pyruvate pH not specified in the publication, 30°C, recombinant mutant DELTAAsp168/Asp171 Thermotoga maritima
1.7
-
 pyruvate pH not specified in the publication, 30°C, recombinant mutant DELTAAsp168 Thermotoga maritima
1.7
-
 pyruvate pH not specified in the publication, 30°C, recombinant mutant DELTAAsp168/Asp237 Thermotoga maritima
1.7
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 45°C, recombinant mutant DELTAAsp168/Asp237 Thermotoga maritima
2.1
-
 pyruvate pH not specified in the publication, 45°C, recombinant mutant DELTAAsp168 Thermotoga maritima
2.2
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 45°C, recombinant mutant DELTAAsp168 Thermotoga maritima
2.4
-
 pyruvate pH not specified in the publication, 45°C, recombinant mutant DELTAAsp168/Asp171 Thermotoga maritima
2.6
-
 pyruvate pH not specified in the publication, 45°C, recombinant mutant DELTAAsp168/Asp237 Thermotoga maritima

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
34000
-
 4 * 34000, SDS-PAGE, role of quaternary structure in the TIM-barrel family of enzymes, overview. Unlike other DHDPS enzymes, but like many thermophilic enzymes, Tm-DHDPS has a large number of charged residues at the quaternary interface. Removal of electrostatic interactions disrupts quaternary structure Thermotoga maritima
130000
-
 gel filtration Thermotoga maritima

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Thermotoga maritima
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-aspartate 4-semialdehyde + pyruvate
-
 Thermotoga maritima (S)-2,3-dihydropyridine-2,6-dicarboxylate + 2 H2O
-
 ?
additional information the active site of the monomer is well conserved, with most active-site residues in the same conformation Thermotoga maritima ?
-
 ?

Subunits

Subunits Comment Organism
tetramer 4 * 34000, SDS-PAGE, role of quaternary structure in the TIM-barrel family of enzymes, overview. Unlike other DHDPS enzymes, but like many thermophilic enzymes, Tm-DHDPS has a large number of charged residues at the quaternary interface. Removal of electrostatic interactions disrupts quaternary structure Thermotoga maritima

Synonyms

Synonyms Comment Organism
DHDPS
-
 Thermotoga maritima
More the enzyme belongs to the TIM-barrel family of enzymes Thermotoga maritima

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30 45 assay at 30°C and at 45°C Thermotoga maritima

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
35
-
 pyruvate pH not specified in the publication, 30°C, recombinant mutant DELTAAsp168 Thermotoga maritima
35
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 30°C, recombinant mutant DELTAAsp168 Thermotoga maritima
39
-
 pyruvate pH not specified in the publication, 30°C, recombinant mutant DELTAAsp237 Thermotoga maritima
39
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 30°C, recombinant mutant DELTAAsp237 Thermotoga maritima
52
-
 pyruvate pH not specified in the publication, 30°C, recombinant mutant DELTAAsp168/Asp171 Thermotoga maritima
52
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 30°C, recombinant mutant DELTAAsp168/Asp171 Thermotoga maritima
63
-
 pyruvate pH not specified in the publication, 30°C, recombinant mutant DELTAAsp171/Arg237 Thermotoga maritima
63
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 30°C, recombinant mutant DELTAAsp171/Arg237 Thermotoga maritima
68
-
 pyruvate pH not specified in the publication, 30°C, recombinant mutant DELTAAsp168/Asp237 Thermotoga maritima
68
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 30°C, recombinant mutant DELTAAsp168/Asp237 Thermotoga maritima
88
-
 pyruvate pH not specified in the publication, 45°C, recombinant mutant DELTAAsp237 Thermotoga maritima
88
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 45°C, recombinant mutant DELTAAsp237 Thermotoga maritima
97
-
 pyruvate pH not specified in the publication, 30°C, recombinant mutant DELTAAsp171 Thermotoga maritima
97
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 30°C, recombinant mutant DELTAAsp171 Thermotoga maritima
108
-
 pyruvate pH not specified in the publication, 45°C, recombinant mutant DELTAAsp168/Asp171 Thermotoga maritima
108
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 45°C, recombinant mutant DELTAAsp168/Asp171 Thermotoga maritima
111
-
 pyruvate pH not specified in the publication, 45°C, recombinant mutant DELTAAsp168 Thermotoga maritima
111
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 45°C, recombinant mutant DELTAAsp168 Thermotoga maritima
134
-
 pyruvate pH not specified in the publication, 45°C, recombinant mutant DELTAAsp171/Arg237 Thermotoga maritima
134
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 45°C, recombinant mutant DELTAAsp171/Arg237 Thermotoga maritima
136
-
 pyruvate pH not specified in the publication, 30°C, recombinant wild-type enzyme Thermotoga maritima
136
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 30°C, recombinant wild-type enzyme Thermotoga maritima
141
-
 pyruvate pH not specified in the publication, 45°C, recombinant mutant DELTAAsp168/Asp237 Thermotoga maritima
141
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 45°C, recombinant mutant DELTAAsp168/Asp237 Thermotoga maritima
233
-
 pyruvate pH not specified in the publication, 45°C, recombinant mutant DELTAAsp171 Thermotoga maritima
233
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 45°C, recombinant mutant DELTAAsp171 Thermotoga maritima
465
-
 pyruvate pH not specified in the publication, 45°C, recombinant wild-type enzyme Thermotoga maritima
465
-
 L-aspartate 4-semialdehyde pH not specified in the publication, 45°C, recombinant wild-type enzyme Thermotoga maritima

General Information

General Information Comment Organism
additional information pyruvate binding occurs near the large interface of DHDPS, and is likely, therefore, to stabilize this solvent-accessible face, which favors the formation of a dimer rather than a monomer. For the DELTAAsp168/Arg237 and DELTAAsp168/Asp171 DHDPS variants addition of pyruvate shifts the equilibrium from primarily monomer to favor almost exclusively dimers. On the other hand, for the DELTAAsp168 DHDPS variant, the monomer-tetramer equilibrium shifts from primarily monomer to primarily tetramer on addition of pyruvate Thermotoga maritima