Cloned (Comment) | Organism |
---|---|
the inhA gene was cloned into the pMK1 mycobacterial expression vector under the control of the strong promoter hsp60. The resulting construct is used to transform Mycobacterium bovis BCG Pasteur in order to allow over-production of recombinant His-tagged InhA | Mycobacterium tuberculosis |
Protein Variants | Comment | Organism |
---|---|---|
T266A | phosphoablative mutant with activity similar to wild-type enzyme | Mycobacterium tuberculosis |
T266D | phosphomimetic mutant with strongly reduced activity (31.4% compared to wild-type enzyme), introduction of inhA_T266D fails to complement growth and mycolic acid defects of an inhA-thermosensitive Mycobacterium smegmatis strain, in a similar manner to what is observed following isoniazid treatment | Mycobacterium tuberculosis |
T266E | phosphomimetic mutant strongly reduced activity (29.5% compared to wild-type enzyme), introduction of inhA_T266E fails to complement growth and mycolic acid defects of an inhA-thermosensitive Mycobacterium smegmatis strain, in a similar manner to what is observed following isoniazid treatment | Mycobacterium tuberculosis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0196 | - |
trans-2-dodecenoyl-CoA | pH 7.5, 25°C, mutant enzyme T266D | Mycobacterium tuberculosis | |
0.0203 | - |
trans-2-dodecenoyl-CoA | pH 7.5, 25°C, mutant enzyme T266E | Mycobacterium tuberculosis | |
0.0409 | - |
trans-2-dodecenoyl-CoA | pH 7.5, 25°C, wild-type enzyme | Mycobacterium tuberculosis | |
0.0523 | - |
trans-2-dodecenoyl-CoA | pH 7.5, 25°C, mutant enzyme T266A | Mycobacterium tuberculosis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycobacterium tuberculosis | P9WGR1 | - |
- |
Mycobacterium tuberculosis ATCC 25618 | P9WGR1 | - |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
phosphoprotein | InhA is phosphorylated in vitro by multiple Ser/Thr kinases on residue Thr266.. Activity of InhA is controlled via phosphorylation. Thr266 is the unique kinase phosphoacceptor, both in vitro and in vivo. The physiological relevance of Thr266 phosphorylation is demonstrated using inhA phosphoablative (T266A) or phosphomimetic (T266D/E) mutants. Enoyl reductase activity is severely impaired in the mimetic mutants in vitro, as a consequence of a reduced binding affinity to NADH. Introduction of inhA_T266D/E fails to complement growth and mycolic acid defects of an inhA-thermosensitive Mycobacterium smegmatis strain, in a similar manner to what is observed following isoniazid treatment. Phosphorylation of InhA may represent an unusual mechanism that allows Mycobacterium tuberculosis to regulate its mycolic acid content, thus offering a new approach to future anti-tuberculosis drug development | Mycobacterium tuberculosis |
Purification (Comment) | Organism |
---|---|
- |
Mycobacterium tuberculosis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
trans-2-dodecenoyl-CoA + NADH + H+ | - |
Mycobacterium tuberculosis | dodecanoyl-CoA + NAD+ | - |
? | |
trans-2-dodecenoyl-CoA + NADH + H+ | - |
Mycobacterium tuberculosis ATCC 25618 | dodecanoyl-CoA + NAD+ | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.46 | - |
trans-2-dodecenoyl-CoA | pH 7.5, 25°C, mutant enzyme T266D | Mycobacterium tuberculosis | |
2.49 | - |
trans-2-dodecenoyl-CoA | pH 7.5, 25°C, mutant enzyme T266E | Mycobacterium tuberculosis | |
5.34 | - |
trans-2-dodecenoyl-CoA | pH 7.5, 25°C, wild-type enzyme | Mycobacterium tuberculosis | |
7.12 | - |
trans-2-dodecenoyl-CoA | pH 7.5, 25°C, mutant enzyme T266A | Mycobacterium tuberculosis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | - |
Mycobacterium tuberculosis |
General Information | Comment | Organism |
---|---|---|
metabolism | the enzyme is an essential enzyme of the mycolic acid biosynthetic pathway | Mycobacterium tuberculosis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
74.49 | - |
trans-2-dodecenoyl-CoA | pH 7.5, 25°C, mutant enzyme T266D | Mycobacterium tuberculosis | |
122.7 | - |
trans-2-dodecenoyl-CoA | pH 7.5, 25°C, mutant enzyme T266E | Mycobacterium tuberculosis | |
130.6 | - |
trans-2-dodecenoyl-CoA | pH 7.5, 25°C, wild-type enzyme | Mycobacterium tuberculosis | |
136.13 | - |
trans-2-dodecenoyl-CoA | pH 7.5, 25°C, mutant enzyme T266A | Mycobacterium tuberculosis |