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Literature summary 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)

EC Number Cloned (Comment) Organism
1.1.1.103 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)

EC Number Crystallization (Comment) Organism
1.1.1.103 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

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.103 0.0099
-
 NAD+ pH 7.5, 65°C, recombinant wild-type enzyme Pyrococcus horikoshii
1.1.1.103 0.0118
-
 L-threonine pH 7.5, 65°C, recombinant wild-type enzyme Pyrococcus horikoshii
1.1.1.103 0.291
-
 NAD+ pH 7.5, 65°C, recombinant mutant R204A Pyrococcus horikoshii
1.1.1.103 0.304
-
 NAD+ pH 7.5, 65°C, recombinant mutant E199A Pyrococcus horikoshii
1.1.1.103 1.51
-
 L-threonine pH 7.5, 65°C, recombinant mutant E199A Pyrococcus horikoshii
1.1.1.103 2.5
-
 L-threonine pH 7.5, 65°C, recombinant mutant R204A Pyrococcus horikoshii

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.103 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]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.103 35000
-
 4 * 35000, recombinant enzyme, SDS-PAGE Pyrococcus horikoshii

Natural Substrates/ Products (Substrates)

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

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.103 Pyrococcus horikoshii O58389
-
-

Substrates and Products (Substrate)

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

Subunits

EC Number Subunits Comment Organism
1.1.1.103 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
1.1.1.103 tetramer 4 * 35000, recombinant enzyme, SDS-PAGE Pyrococcus horikoshii

Synonyms

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.103 65
-
 assay at Pyrococcus horikoshii

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.103 0.55
-
 NAD+ pH 7.5, 65°C, recombinant mutant E199A Pyrococcus horikoshii
1.1.1.103 0.55
-
 L-threonine pH 7.5, 65°C, recombinant mutant E199A Pyrococcus horikoshii
1.1.1.103 0.667
-
 NAD+ pH 7.5, 65°C, recombinant mutant R204A Pyrococcus horikoshii
1.1.1.103 0.667
-
 L-threonine pH 7.5, 65°C, recombinant mutant R204A Pyrococcus horikoshii
1.1.1.103 11
-
 NAD+ pH 7.5, 65°C, recombinant wild-type enzyme Pyrococcus horikoshii
1.1.1.103 11
-
 L-threonine pH 7.5, 65°C, recombinant wild-type enzyme Pyrococcus horikoshii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.103 7.5
-
 assay at Pyrococcus horikoshii

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.103 NAD+
-
 Pyrococcus horikoshii
1.1.1.103 NADH
-
 Pyrococcus horikoshii