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Literature summary for 1.1.1.103 extracted from

  • Ishikawa, K.; Higashi, N.; Nakamura, T.; Matsuura, T.; Nakagawa, A.
    The first crystal structure of L-threonine dehydrogenase (2007), J. Mol. Biol., 366, 857-867.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
orf PH0655, NA and amino acid sequence determination, analysis, and comparison, expression of selenomethionine-labeled wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Pyrococcus horikoshii

Crystallization (Commentary)

Crystallization (Comment) Organism
selenomethionine-substituted enzyme, 10 mg/ml protein in 50 mM Tris-HCl buffer, pH 7.5, hanging-drop vapor-diffusion method at 4°C, mixing of 0.0015 ml of each, protein and reservoir solution, the latter containing 0.2 M sodium chloride, 0.1 M HEPES buffer, pH 7.5, and 40% v/v PEG 400, five days, X-ray diffraction structure determination and analysis, single wavelength anomalous diffraction method Pyrococcus horikoshii

Protein Variants

Protein Variants Comment Organism
E199A site-directed mutagenesis, the mutant shows altered kinetics compared to the wild-type enzyme Pyrococcus horikoshii
R204A site-directed mutagenesis, the mutant shows altered kinetics compared to the wild-type enzyme Pyrococcus horikoshii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0099
-
NAD+ pH 7.5, 65°C, recombinant wild-type enzyme Pyrococcus horikoshii
0.0118
-
L-threonine pH 7.5, 65°C, recombinant wild-type enzyme Pyrococcus horikoshii
0.291
-
NAD+ pH 7.5, 65°C, recombinant mutant R204A Pyrococcus horikoshii
0.304
-
NAD+ pH 7.5, 65°C, recombinant mutant E199A Pyrococcus horikoshii
1.51
-
L-threonine pH 7.5, 65°C, recombinant mutant E199A Pyrococcus horikoshii
2.5
-
L-threonine pH 7.5, 65°C, recombinant mutant R204A Pyrococcus horikoshii

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+ one molecule of TDH contains one zinc ion playing a structural role, the metal ion is ligated by foru Cys residues, coenzyme-binding domain shows a larger interdomain cleft compared to other ADHs, binding structure, overview Pyrococcus horikoshii

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
35000
-
4 * 35000, recombinant enzyme, SDS-PAGE Pyrococcus horikoshii

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-threonine + NAD+ Pyrococcus horikoshii
-
(2S)-2-amino-3-oxobutanoate + NADH + H+
-
r

Organism

Organism UniProt Comment Textmining
Pyrococcus horikoshii O58389
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-threonine + NAD+
-
Pyrococcus horikoshii (2S)-2-amino-3-oxobutanoate + NADH + H+
-
r

Subunits

Subunits Comment Organism
More crystal structure analysis, each subunit is composed of two domains: an NAD(H)-binding domain and a catalytic domain. The NAD(H)-binding domain contains the alpha/beta Rossmann fold motif, characteristic of the NAD(H)-binding protein Pyrococcus horikoshii
tetramer 4 * 35000, recombinant enzyme, SDS-PAGE Pyrococcus horikoshii

Synonyms

Synonyms Comment Organism
L-threonine dehydrogenase
-
Pyrococcus horikoshii
More the enzyme belongs to the medium-chain NAD(H)-dependent alcohol dehydrogenases Pyrococcus horikoshii
TDH
-
Pyrococcus horikoshii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
65
-
assay at Pyrococcus horikoshii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.55
-
NAD+ pH 7.5, 65°C, recombinant mutant E199A Pyrococcus horikoshii
0.55
-
L-threonine pH 7.5, 65°C, recombinant mutant E199A Pyrococcus horikoshii
0.667
-
NAD+ pH 7.5, 65°C, recombinant mutant R204A Pyrococcus horikoshii
0.667
-
L-threonine pH 7.5, 65°C, recombinant mutant R204A Pyrococcus horikoshii
11
-
NAD+ pH 7.5, 65°C, recombinant wild-type enzyme Pyrococcus horikoshii
11
-
L-threonine pH 7.5, 65°C, recombinant wild-type enzyme Pyrococcus horikoshii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Pyrococcus horikoshii

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Pyrococcus horikoshii
NADH
-
Pyrococcus horikoshii