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Literature summary for 1.1.1.103 extracted from

  • Johnson, A.R.; Chen, Y.W.; Dekker, E.E.
    Investigation of a catalytic zinc binding site in Escherichia coli L-threonine dehydrogenase by site-directed mutagenesis of cysteine-38 (1998), Arch. Biochem. Biophys., 358, 211-221.
    View publication on PubMed

Protein Variants

Protein Variants Comment Organism
C38D site-directed mutagenesis Escherichia coli K-12
C38S site-directed mutagenesis, catalytically inactive mutant, C38 is located in an activating divalent metal-ion binding site Escherichia coli K-12

Inhibitors

Inhibitors Comment Organism Structure
Hg2+ 0.5 mM, 100% inhibition of wild-type and C38D mutant enzyme Escherichia coli K-12
Mn2+ preincubation with 1 mM, 47% inhibition of L-threonine amide oxidation, 73% inhibition of L-threonine methyl ester oxidation, 59% inhibition of L-serine oxidation, 48% inhibition of D,L-threo-beta-phenylserine oxidation Escherichia coli K-12

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.75
-
NAD+ C38D mutant Escherichia coli K-12

Metals/Ions

Metals/Ions Comment Organism Structure
Cd2+ preincubation with 0.07 or 5 mM leads to 3fold increase of C38D mutant enzyme activity, 0.5 mM, 6.5fold increase of wild-type enzyme activity, substrate L-threonine Escherichia coli K-12
Co2+ 5 mM, 6fold increase of C38D mutant enzyme activity Escherichia coli K-12
Mn2+ preincubation with 1 mM, 3.5fold increase of L-threonine oxidation Escherichia coli K-12
Zn2+ 5 mM, 60fold increase of C38D mutant enzyme activity, Zn2+ stimulated activity decreases to 10% when 1 mM Mn2+ is added, no increase in activity for wild-type enzyme Escherichia coli K-12

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
149000
-
non-denaturing PAGE, wild-type and C38D mutant Escherichia coli K-12

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-threonine + NAD+ Escherichia coli K-12
-
(2S)-2-amino-3-oxobutanoate + NADH + H+
-
r

Organism

Organism UniProt Comment Textmining
Escherichia coli K-12
-
-
-

Purification (Commentary)

Purification (Comment) Organism
treatment of cell extract at 60°C, DEAE-Sephadex, Mn2+ activation, Blue 2-Sepharose CL-6B, C38D mutant was purified without Mn2+ activation Escherichia coli K-12

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.1
-
C38D mutant, incubated with 1 mM Mn2+ 15 min before assay Escherichia coli K-12
0.2
-
C38D mutant Escherichia coli K-12
0.6
-
C38D mutant, incubated with 0.07 or 5 mM Cd2+ 15 min before assay Escherichia coli K-12
1.2
-
C38D mutant, incubated with 5 mM Co2+ 15 min before assay Escherichia coli K-12
2
-
C38D mutant, incubated with 5 mM Zn2+ and 1 mM Mn2+ 15 min before assay Escherichia coli K-12
10.2
-
C38D mutant, incubated with 5 mM Zn2+ and 0.07 mM Cd2+ 15 min before assay Escherichia coli K-12
11.4
-
C38D mutant, incubated with 5 mM Zn2+ 15 min before assay Escherichia coli K-12

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
DL-2-amino-3-hydroxypentanoate + NAD+
-
Escherichia coli K-12 L-2-amino-3-oxopentanoate + NADH
-
?
DL-threo-beta-phenylserine + NAD+
-
Escherichia coli K-12 DL-2-amino-3-phenyl-3-oxopropionate + NADH
-
?
L-serine + NAD+
-
Escherichia coli K-12 L-2-amino-3-oxopropionate + NADH + H+
-
?
L-threonine + NAD+
-
Escherichia coli K-12 (2S)-2-amino-3-oxobutanoate + NADH + H+
-
r
L-threonine amide + NAD+
-
Escherichia coli K-12 L-2-amino-3-oxobutyramide + NADH
-
?
L-threonine methyl ester + NAD+
-
Escherichia coli K-12 L-2-amino-3-oxobutanoate methyl ester + NADH
-
?

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
C38D mutant enzyme Escherichia coli K-12
8.4
-
C38D mutant enzyme, stimulated with 5 mM Zn2+ Escherichia coli K-12
10.1
-
wild-type enzyme Escherichia coli K-12

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Escherichia coli K-12