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Literature summary extracted from

  • Soares da Costa, T.P.; Muscroft-Taylor, A.C.; Dobson, R.C.; Devenish, S.R.; Jameson, G.B.; Gerrard, J.A.
    How essential is the essential active-site lysine in dihydrodipicolinate synthase? (2010), Biochimie, 92, 837-845.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.3.3.7 DHDPS overexpressed in Escherichia coli AT997recA-, transformed with site-directed mutants based on the pBluescript plasmid pJG001 Escherichia coli

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
4.3.3.7 by the hanging drop-vapour diffusion method, mutants K161A and K161R solved at resolutions of 2.0 and 2.1 A, respectively. They show no changes in their secondary or tertiary structures when compared to the wild-type structure. Crystal structure of mutant K161A with pyruvate bound at the active site solved at a resolution of 2.3 A, reveals a defined binding pocket for pyruvate that is thus not dependent upon lysine 161 Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
4.3.3.7 K161A catalytically active, significant decrease in activity. Is not inactivated when incubated with pyruvate and the reducing agent sodium borohydride. Negligible heat production associated with pyruvate binding to the mutant enzyme, consistent with the lack of Schiff base formation Escherichia coli
4.3.3.7 K161R catalytically active, significant decrease in activity. Is not inactivated when incubated with pyruvate and the reducing agent sodium borohydride. Negligible heat production associated with pyruvate binding to the mutant enzyme, consistent with the lack of Schiff base formation Escherichia coli

Inhibitors

EC Number Inhibitors Comment Organism Structure
4.3.3.7 lysine inhibition of wild-type DHDPS by lysine with respect to pyruvate is partial and uncompetitive, and partial non-competitive with respect to L-aspartate 4-semialdehyde. Ethanolamine, n-butylamine, 1-amino-2-propanol, 3-amino-1-propanol, iso-butylamine and Tris-HCl cannot rescue activity Escherichia coli
4.3.3.7 Sodium borohydride wild-type DHDPS is inactivated when incubated with pyruvate, whereas incubation with L-aspartate 4-semialdehyde has no effect Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.3.3.7 0.12
-
 L-aspartate 4-semialdehyde mutant K161R, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR Escherichia coli
4.3.3.7 0.12
-
 L-aspartate 4-semialdehyde wild-type, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR Escherichia coli
4.3.3.7 0.15
-
 pyruvate wild-type, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR Escherichia coli
4.3.3.7 0.23
-
 L-aspartate 4-semialdehyde mutant K161A, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR Escherichia coli
4.3.3.7 0.45
-
 pyruvate mutant K161A, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR Escherichia coli
4.3.3.7 0.57
-
 pyruvate mutant K161R, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR Escherichia coli

Organism

EC Number Organism UniProt Comment Textmining
4.3.3.7 Escherichia coli P0A6L2
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.3.3.7 wild-type and mutants, by gel filtration Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.3.3.7 L-aspartate 4-semialdehyde + pyruvate condensation reaction between both substrates via the formation of a Schiff base intermediate between pyruvate and the absolutely conserved active-site lysine. Although lysine 161 is important in the wild-type DHDPS-catalysed reaction, it is not absolutely essential for catalysis Escherichia coli dihydrodipicolinate + 2 H2O
-
 ?

Synonyms

EC Number Synonyms Comment Organism
4.3.3.7 DHDPS
-
 Escherichia coli
4.3.3.7 dihydrodipicolinate synthase
-
 Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.3.3.7 0.06
-
 pyruvate mutant K161A, at 30°C, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR Escherichia coli
4.3.3.7 0.16
-
 pyruvate mutant K161R, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR Escherichia coli
4.3.3.7 45
-
 pyruvate wild-type, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR Escherichia coli

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
4.3.3.7 0.12
-
 lysine wild-type, with pyruvate as substrate Escherichia coli
4.3.3.7 0.14
-
 lysine mutant K161A, with pyruvate as substrate Escherichia coli
4.3.3.7 0.14
-
 lysine mutant K161R, with L-aspartate 4-semialdehyde as substrate Escherichia coli
4.3.3.7 0.14
-
 lysine mutant K161R, with pyruvate as substrate Escherichia coli
4.3.3.7 0.18
-
 lysine wild-type, with L-aspartate 4-semialdehyde as substrate Escherichia coli
4.3.3.7 0.23
-
 lysine mutant K161A, with L-aspartate 4-semialdehyde as substrate Escherichia coli

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4.3.3.7 0.13
-
 pyruvate mutant K161A, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR Escherichia coli
4.3.3.7 0.26
-
 L-aspartate 4-semialdehyde mutant K161A, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR Escherichia coli
4.3.3.7 0.28
-
 pyruvate mutant K161R, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR Escherichia coli
4.3.3.7 1.3
-
 L-aspartate 4-semialdehyde mutant K161R, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR Escherichia coli
4.3.3.7 300
-
 pyruvate wild-type, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR Escherichia coli
4.3.3.7 380
-
 L-aspartate 4-semialdehyde wild-type, in 100 mM HEPES buffer, pH 8.0, 0.2 mM NADPH, 50 microg/ml DHDPR Escherichia coli