Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(lambdaDE3) cells | Mycobacterium tuberculosis |
Crystallization (Comment) | Organism |
---|---|
native enzyme, the holoenzyme, binary complexes with NAD+, (S)-3-hydroxyisobutyrate, (R)-3-hydroxyisobutyrate, L-serine and 3-hydroxypropanoic acid and ternary complexes involving the substrates and NAD+, microbatch-under-oil method, using 0.02 M MgCl2, 0.1 M HEPES (pH 7.5), 22% (w/v) polyacrylic acid sodium salt 5100 | Mycobacterium tuberculosis |
Protein Variants | Comment | Organism |
---|---|---|
D244A | the mutant shows strong reduction in the activity compared to the wild type enzyme | Mycobacterium tuberculosis |
D275A | the mutant shows a marginal loss of activity compared to the wild type enzyme | Mycobacterium tuberculosis |
F236A | the mutant shows reduction in the activity compared to the wild type enzyme | Mycobacterium tuberculosis |
K168A | inactive | Mycobacterium tuberculosis |
N172A | the mutant shows dramatic reduction in the activity compared to the wild type enzyme | Mycobacterium tuberculosis |
S119A | inactive | Mycobacterium tuberculosis |
T207A | the mutant shows reduction in the activity compared to the wild type enzyme | Mycobacterium tuberculosis |
W211A | the mutant shows dramatic reduction in the activity compared to the wild type enzyme | Mycobacterium tuberculosis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
NADH | competitive inhibition in the presence of NADH when NAD+ is varied, noncompetitive inhibition with NADH when (S)-3-hydroxyisobutyrate is varied | Mycobacterium tuberculosis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.563 | - |
NAD+ | with (S)-3-hydroxyisobutyrate as cosubstrate, pH 8.5 and 30°C | Mycobacterium tuberculosis | |
0.574 | - |
(S)-3-hydroxyisobutyrate | at pH 8.5 and 30°C | Mycobacterium tuberculosis | |
0.582 | - |
NADP+ | with (S)-3-hydroxyisobutyrate as cosubstrate, pH 8.5 and 30°C | Mycobacterium tuberculosis | |
2.5 | 3 | 3-hydroxypropanoic acid | at pH 8.5 and 30°C | Mycobacterium tuberculosis | |
4.56 | - |
(R)-3-hydroxyisobutyrate | at pH 8.5 and 30°C | Mycobacterium tuberculosis | |
21.33 | - |
L-serine | at pH 8.5 and 30°C | Mycobacterium tuberculosis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
66000 | - |
gel filtration | Mycobacterium tuberculosis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-3-hydroxyisobutyrate + NAD+ | Mycobacterium tuberculosis | the enzyme is specific to (S)-3-hydroxyisobutyrate and prefers NAD+ as cofactor | 2-methyl-3-oxopropanoate + NADH + H+ | - |
? | |
(S)-3-hydroxyisobutyrate + NAD+ | Mycobacterium tuberculosis H37Rv | the enzyme is specific to (S)-3-hydroxyisobutyrate and prefers NAD+ as cofactor | 2-methyl-3-oxopropanoate + NADH + H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycobacterium tuberculosis | P9WNY5 | - |
- |
Mycobacterium tuberculosis H37Rv | P9WNY5 | - |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA resin column chromatography | Mycobacterium tuberculosis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R)-3-hydroxyisobutyrate + NAD+ | substrate with much less efficiency compared to (S)-3-hydroxyisobutyrate | Mycobacterium tuberculosis | 2-methyl-3-oxopropanoate + NADH + H+ | - |
? | |
(R)-3-hydroxyisobutyrate + NAD+ | substrate with much less efficiency compared to (S)-3-hydroxyisobutyrate | Mycobacterium tuberculosis H37Rv | 2-methyl-3-oxopropanoate + NADH + H+ | - |
? | |
(S)-3-hydroxyisobutyrate + NAD+ | the enzyme is specific to (S)-3-hydroxyisobutyrate and prefers NAD+ as cofactor | Mycobacterium tuberculosis | 2-methyl-3-oxopropanoate + NADH + H+ | - |
? | |
(S)-3-hydroxyisobutyrate + NAD+ | the enzyme is specific to (S)-3-hydroxyisobutyrate | Mycobacterium tuberculosis | 2-methyl-3-oxopropanoate + NADH + H+ | - |
? | |
(S)-3-hydroxyisobutyrate + NAD+ | the enzyme is specific to (S)-3-hydroxyisobutyrate and prefers NAD+ as cofactor | Mycobacterium tuberculosis H37Rv | 2-methyl-3-oxopropanoate + NADH + H+ | - |
? | |
(S)-3-hydroxyisobutyrate + NAD+ | the enzyme is specific to (S)-3-hydroxyisobutyrate | Mycobacterium tuberculosis H37Rv | 2-methyl-3-oxopropanoate + NADH + H+ | - |
? | |
(S)-3-hydroxyisobutyrate + NADP+ | - |
Mycobacterium tuberculosis | 2-methyl-3-oxopropanoate + NADPH + H+ | - |
? | |
3-hydroxypropanoic acid + NAD+ | substrate with much less efficiency compared to (S)-3-hydroxyisobutyrate | Mycobacterium tuberculosis | 3-oxopropanoate + NADH + H+ | - |
? | |
L-serine + NAD+ | substrate with much less efficiency compared to (S)-3-hydroxyisobutyrate | Mycobacterium tuberculosis | ? | - |
? | |
L-serine + NAD+ | substrate with much less efficiency compared to (S)-3-hydroxyisobutyrate | Mycobacterium tuberculosis H37Rv | ? | - |
? | |
additional information | no activity with D-serine, (S)-3-hydroxyisobutyrate methyl ester, (R)-3-hydroxyisobutyrate methyl ester, L-threonine and D-threonine | Mycobacterium tuberculosis | ? | - |
- |
|
additional information | no activity with D-serine, (S)-3-hydroxyisobutyrate methyl ester, (R)-3-hydroxyisobutyrate methyl ester, L-threonine and D-threonine | Mycobacterium tuberculosis H37Rv | ? | - |
- |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 30875, calculated from amino acid sequence | Mycobacterium tuberculosis |
Synonyms | Comment | Organism |
---|---|---|
3-hydroxyisobutyric acid dehydrogenase | - |
Mycobacterium tuberculosis |
HIBADH | - |
Mycobacterium tuberculosis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
34.5 | - |
at pH 11.0, the melting temperature is 34.5°C | Mycobacterium tuberculosis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.205 | - |
(R)-3-hydroxyisobutyrate | at pH 8.5 and 30°C | Mycobacterium tuberculosis | |
0.262 | - |
L-serine | at pH 8.5 and 30°C | Mycobacterium tuberculosis | |
0.29 | - |
NADP+ | with (S)-3-hydroxyisobutyrate as cosubstrate, pH 8.5 and 30°C | Mycobacterium tuberculosis | |
1.018 | - |
(S)-3-hydroxyisobutyrate | at pH 8.5 and 30°C | Mycobacterium tuberculosis | |
1.182 | - |
NAD+ | with (S)-3-hydroxyisobutyrate as cosubstrate, pH 8.5 and 30°C | Mycobacterium tuberculosis | |
2.53 | - |
3-hydroxypropanoic acid | at pH 8.5 and 30°C | Mycobacterium tuberculosis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
11 | - |
- |
Mycobacterium tuberculosis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | the enzyme preferentially uses NAD+ as the cofactor | Mycobacterium tuberculosis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.044 | - |
L-serine | at pH 8.5 and 30°C | Mycobacterium tuberculosis | |
0.0448 | - |
(R)-3-hydroxyisobutyrate | at pH 8.5 and 30°C | Mycobacterium tuberculosis | |
0.1036 | - |
3-hydroxypropanoic acid | at pH 8.5 and 30°C | Mycobacterium tuberculosis | |
0.498 | - |
NADP+ | with (S)-3-hydroxyisobutyrate as cosubstrate, pH 8.5 and 30°C | Mycobacterium tuberculosis | |
1.773 | - |
(S)-3-hydroxyisobutyrate | at pH 8.5 and 30°C | Mycobacterium tuberculosis | |
2.1 | - |
NAD+ | with (S)-3-hydroxyisobutyrate as cosubstrate, pH 8.5 and 30°C | Mycobacterium tuberculosis |