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The C-terminal domain of Escherichia coli dihydrodipicolinate synthase (DHDPS) is essential for maintenance of quaternary structure and efficient catalysis

Guo, B.B.; Devenish, S.R.; Dobson, R.C.; Muscroft-Taylor, A.C.; Gerrard, J.A.; Biochem. Biophys. Res. Commun. 380, 802-806 (2009)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
4.3.3.7
the dapA and dapA-H225* genes introduced into the vector pUCX, to transform the auxotrophic Escherichia coli XL1-Blue KanRDELTAdapA cells, under the control of a lac promoter/repressor system. Mutations in dapA introduced into the pET-151/D-TOPO plasmid. C-terminal truncated DHDPS (H225*) expressed in Escherichia coli BL21 Star (DE3)
Escherichia coli
Engineering
EC Number
Amino acid exchange
Commentary
Organism
4.3.3.7
additional information
C-terminal truncated DHDPS (H225*), exhibits a dramatic reduction in both solubility and stability. Equilibrating mixture of quaternary states. The substrate, pyruvate, and the feedback inhibitor, (S)-lysine, both have a positive effect on thermostability of C-terminal truncated DHDPS (H225*)
Escherichia coli
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
4.3.3.7
L-lysine
-
Escherichia coli
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.3.3.7
0.23
-
L-aspartate 4-semialdehyde
C-terminal truncated DHDPS (H225*), at 30°C, in 150 mM HEPES, pH 8.0, 0.16 mM NADPH, 50 microg/ml DHDPR
Escherichia coli
4.3.3.7
0.29
-
L-aspartate 4-semialdehyde
polyhistidine-tagged wild-type DHDPSR
Escherichia coli
4.3.3.7
0.36
-
pyruvate
polyhistidine-tagged wild-type DHDPS
Escherichia coli
4.3.3.7
0.37
-
pyruvate
C-terminal truncated DHDPS (H225*), at 30°C, in 150 mM HEPES, pH 8.0, 0.16 mM NADPH, 50 microg/ml DHDPR
Escherichia coli
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
4.3.3.7
Escherichia coli
P0A6L2
-
-
Purification (Commentary)
EC Number
Commentary
Organism
4.3.3.7
-
Escherichia coli
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4.3.3.7
L-aspartate 4-semialdehyde + pyruvate
-
702036
Escherichia coli
dihydrodipicolinate + 2 H2O
-
-
-
?
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
4.3.3.7
1.2
-
pyruvate
C-terminal truncated DHDPS (H225*), at 30°C, in 150 mM HEPES, pH 8.0, 0.16 mM NADPH, 50 microg/ml DHDPR
Escherichia coli
4.3.3.7
75
-
pyruvate
polyhistidine-tagged wild-type DHDPS
Escherichia coli
IC50 Value
EC Number
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
4.3.3.7
0.21
-
wild-type DHDPS
Escherichia coli
L-lysine
4.3.3.7
0.22
-
C-terminal truncated DHDPS (H225*)
Escherichia coli
L-lysine
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
4.3.3.7
the dapA and dapA-H225* genes introduced into the vector pUCX, to transform the auxotrophic Escherichia coli XL1-Blue KanRDELTAdapA cells, under the control of a lac promoter/repressor system. Mutations in dapA introduced into the pET-151/D-TOPO plasmid. C-terminal truncated DHDPS (H225*) expressed in Escherichia coli BL21 Star (DE3)
Escherichia coli
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
4.3.3.7
additional information
C-terminal truncated DHDPS (H225*), exhibits a dramatic reduction in both solubility and stability. Equilibrating mixture of quaternary states. The substrate, pyruvate, and the feedback inhibitor, (S)-lysine, both have a positive effect on thermostability of C-terminal truncated DHDPS (H225*)
Escherichia coli
IC50 Value (protein specific)
EC Number
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
4.3.3.7
0.21
-
wild-type DHDPS
Escherichia coli
L-lysine
4.3.3.7
0.22
-
C-terminal truncated DHDPS (H225*)
Escherichia coli
L-lysine
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
4.3.3.7
L-lysine
-
Escherichia coli
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.3.3.7
0.23
-
L-aspartate 4-semialdehyde
C-terminal truncated DHDPS (H225*), at 30°C, in 150 mM HEPES, pH 8.0, 0.16 mM NADPH, 50 microg/ml DHDPR
Escherichia coli
4.3.3.7
0.29
-
L-aspartate 4-semialdehyde
polyhistidine-tagged wild-type DHDPSR
Escherichia coli
4.3.3.7
0.36
-
pyruvate
polyhistidine-tagged wild-type DHDPS
Escherichia coli
4.3.3.7
0.37
-
pyruvate
C-terminal truncated DHDPS (H225*), at 30°C, in 150 mM HEPES, pH 8.0, 0.16 mM NADPH, 50 microg/ml DHDPR
Escherichia coli
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
4.3.3.7
-
Escherichia coli
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4.3.3.7
L-aspartate 4-semialdehyde + pyruvate
-
702036
Escherichia coli
dihydrodipicolinate + 2 H2O
-
-
-
?
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
4.3.3.7
1.2
-
pyruvate
C-terminal truncated DHDPS (H225*), at 30°C, in 150 mM HEPES, pH 8.0, 0.16 mM NADPH, 50 microg/ml DHDPR
Escherichia coli
4.3.3.7
75
-
pyruvate
polyhistidine-tagged wild-type DHDPS
Escherichia coli
General Information
EC Number
General Information
Commentary
Organism
4.3.3.7
physiological function
complementation of the auxotrophy of Escherichia coli XL1-Blue KanRDELTAdapA cells only with the plasmid pUCX:dapA encoding wild-type DHDPS
Escherichia coli
General Information (protein specific)
EC Number
General Information
Commentary
Organism
4.3.3.7
physiological function
complementation of the auxotrophy of Escherichia coli XL1-Blue KanRDELTAdapA cells only with the plasmid pUCX:dapA encoding wild-type DHDPS
Escherichia coli