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 | 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 Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.75 | - |
NAD+ | C38D mutant | Escherichia coli K-12 |
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 Maximum [Da] | Comment | Organism |
---|---|---|---|
149000 | - |
non-denaturing PAGE, wild-type and C38D mutant | Escherichia coli K-12 |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli K-12 | - |
- |
- |
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 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 | 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 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 | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Escherichia coli K-12 |