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Literature summary extracted from

  • Yew, W.S.; Fedorov, A.A.; Fedorov, E.V.; Wood, B.M.; Almo, S.C.; Gerlt, J.A.
    Evolution of enzymatic activities in the enolase superfamily: D-tartrate dehydratase from Bradyrhizobium japonicum (2006), Biochemistry, 45, 14598-14608.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.1.81 expression in Escherichia coli strain BL21 Bradyrhizobium japonicum

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
4.2.1.81 all crystals, wild type TarD liganded with Mg2+, selenomethionine-substituted TarD liganded with Mg2+, wild-type TarD liganded with Mg2+ and meso-tartrate, and the K184A mutant liganded with Mg2+ and D-tartrate, are grown by the hanging drop method at room temperature Bradyrhizobium japonicum

Protein Variants

EC Number Protein Variants Comment Organism
4.2.1.81 H322A kcatKm is 193000fold lower than wild-type value Bradyrhizobium japonicum
4.2.1.81 H322N kcatKm is 607fold lower than wild-type value Bradyrhizobium japonicum
4.2.1.81 H322Q kcatKm is 340000fold lower than wild-type value Bradyrhizobium japonicum
4.2.1.81 K102A inactive mutant protein Bradyrhizobium japonicum
4.2.1.81 K102M inactive mutant protein Bradyrhizobium japonicum
4.2.1.81 K184A inactive mutant enzyme Bradyrhizobium japonicum
4.2.1.81 K184R reduced activity. kcat/Km is 500fold lower than wild-type value Bradyrhizobium japonicum

Inhibitors

EC Number Inhibitors Comment Organism Structure
4.2.1.81 meso-tartrate competitive inhibitor Bradyrhizobium japonicum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.2.1.81 0.035
-
 (S,S)-Tartrate 25°C, pH 7.5, mutant enzyme K184R Bradyrhizobium japonicum
4.2.1.81 0.048
-
 (S,S)-Tartrate 25°C, pH 7.5, mutant enzyme H322N Bradyrhizobium japonicum
4.2.1.81 0.086
-
 (S,S)-Tartrate 25°C, pH 7.5, wild-type enzyme Bradyrhizobium japonicum
4.2.1.81 6.2
-
 (S,S)-Tartrate 25°C, pH 7.5, mutant enzyme H322A Bradyrhizobium japonicum
4.2.1.81 7.7
-
 (S,S)-Tartrate 25°C, pH 7.5, mutant enzyme H322Q Bradyrhizobium japonicum

Organism

EC Number Organism UniProt Comment Textmining
4.2.1.81 Bradyrhizobium japonicum
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.1.81
-
 Bradyrhizobium japonicum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.81 (S,S)-tartrate mechanism in which Lys 184 initiates the reaction by abstraction of the R-proton to generate a Mg2+-stabilized enediolate intermediate, and the vinylogous beta-elimination of the 3-OH group is general acid-catalyzed by the His 322, accomplishing the anti-elimination of water. The replacement of the leaving group by solvent-derived hydrogen is stereorandom Bradyrhizobium japonicum oxaloacetate + H2O
-
 ?

Synonyms

EC Number Synonyms Comment Organism
4.2.1.81 bll6730
-
 Bradyrhizobium japonicum
4.2.1.81 TarD
-
 Bradyrhizobium japonicum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.2.1.81 0.0019
-
 (S,S)-Tartrate 25°C, pH 7.5, mutant enzyme H322Q Bradyrhizobium japonicum
4.2.1.81 0.0027
-
 (S,S)-Tartrate 25°C, pH 7.5, mutant enzyme H322A Bradyrhizobium japonicum
4.2.1.81 0.0058
-
 (S,S)-Tartrate 25°C, pH 7.5, mutant enzyme K184R Bradyrhizobium japonicum
4.2.1.81 0.0069
-
 (S,S)-Tartrate 25°C, pH 7.5, mutant enzyme H322N Bradyrhizobium japonicum
4.2.1.81 7.3
-
 (S,S)-Tartrate 25°C, pH 7.5, wild-type enzyme Bradyrhizobium japonicum