Application | Comment | Organism |
---|---|---|
medicine | TBNAT remains a potential drug target for the treatment of tuberculosis | Mycobacterium tuberculosis |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli as a His-tagged fusion protein | Mycobacterium tuberculosis |
into the pET28a+ vector for expression in Escherichia coli BL21DE3 cells | Mycobacterium tuberculosis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
iodoacetamide | incubation of TBNAT with iodoacetamide results in a time-dependentloss of enzymatic activity | Mycobacterium tuberculosis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.32 | - |
3-Amino-4-hydroxybenzoic acid | - |
Mycobacterium tuberculosis | |
0.61 | - |
hydralazine | - |
Mycobacterium tuberculosis | |
0.9 | - |
4-amino-3-hydroxybenzoic acid | - |
Mycobacterium tuberculosis | |
1.9 | - |
2-amino-4-chlorophenol | - |
Mycobacterium tuberculosis | |
2 | 3 | aniline | - |
Mycobacterium tuberculosis | |
3.06 | - |
2-amino-4-methylphenol | - |
Mycobacterium tuberculosis | |
5 | - |
p-Aminobenzoic acid | - |
Mycobacterium tuberculosis | |
5.79 | - |
2-Aminophenol | - |
Mycobacterium tuberculosis | |
6 | - |
4-chloroaniline | - |
Mycobacterium tuberculosis | |
11 | - |
4-methylaniline | - |
Mycobacterium tuberculosis | |
14 | - |
anisidine | - |
Mycobacterium tuberculosis | |
14 | - |
nicotinic acid hydrazide | - |
Mycobacterium tuberculosis | |
14 | - |
benzoic acid hydrazide | - |
Mycobacterium tuberculosis | |
20 | - |
4-hydroxybenzhydrazide | - |
Mycobacterium tuberculosis | |
51 | - |
4-fluoroaniline | - |
Mycobacterium tuberculosis | |
102 | - |
isoniazid | - |
Mycobacterium tuberculosis |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytosol | - |
Mycobacterium tuberculosis | 5829 | - |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
30000 | - |
SDS-PAGE | Mycobacterium tuberculosis |
30000 | - |
determined by SDS-PAGE | Mycobacterium tuberculosis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycobacterium tuberculosis | - |
- |
- |
Purification (Comment) | Organism |
---|---|
using a Ni-NTA and a Superdex S-75 column | Mycobacterium tuberculosis |
using Ni-NTA chromatography. Approximately 2.7 mg of purified enzyme is obtained per liter of culture | Mycobacterium tuberculosis |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
acetyl-CoA + an arylamine = CoA + an N-acetylarylamine | steady-state kinetic analysis of TBNAT reveals that the enzyme catalyzes the reaction via a bi-bi ping-pong kinetic mechanism: enzyme initially reacts with AcCoA to generate the first product, CoA, and the acetyl-enzyme intermediate (E-Ac). The amine binds and reacts with E-Ac to generate the second product, acetylated amine, and regenerate the free enzyme. The pH dependence of the kinetic parameters reveals that one enzyme group must be deprotonated for optimal catalytic activity and that two amino acid residues at the active site of the free enzyme are involved in binding and/or catalysis. Solvent kinetic isotope effects suggest that proton transfer steps are not rate-limiting in the overall reaction for substituted aniline substrates but become rate-limiting when poor hydrazide substrates are used | Mycobacterium tuberculosis |
Storage Stability | Organism |
---|---|
-20°C, 50% glycerol | Mycobacterium tuberculosis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-CoA + 2-amino-4-chlorophenol | - |
Mycobacterium tuberculosis | CoA + N-(5-chloro-2-hydroxyphenyl)acetamide | - |
? | |
acetyl-CoA + 2-amino-4-chlorophenol | - |
Mycobacterium tuberculosis | CoA + N-acetyl-2-amino-4-chlorophenol | - |
? | |
acetyl-CoA + 2-amino-4-methylphenol | - |
Mycobacterium tuberculosis | CoA + N-(2-hydroxy-5-methylphenyl)acetamide | - |
? | |
acetyl-CoA + 2-amino-4-methylphenol | - |
Mycobacterium tuberculosis | CoA + N-acetyl-2-amino-4-methylphenol | - |
? | |
acetyl-CoA + 2-aminophenol | - |
Mycobacterium tuberculosis | CoA + N-(2-hydroxyphenyl)acetamide | - |
? | |
acetyl-CoA + 2-aminophenol | - |
Mycobacterium tuberculosis | CoA + N-acetyl-2-aminophenol | - |
? | |
acetyl-CoA + 3-amino-4-hydroxybenzoic acid | - |
Mycobacterium tuberculosis | CoA + 3-(acetylamino)-4-hydroxybenzoic acid | - |
? | |
acetyl-CoA + 3-amino-4-hydroxybenzoic acid | - |
Mycobacterium tuberculosis | CoA + N-acetyl-3-amino-4-hydroxybenzoic acid | - |
? | |
acetyl-CoA + 4-amino-3-hydroxybenzoic acid | - |
Mycobacterium tuberculosis | CoA + 4-(acetylamino)-3-hydroxybenzoic acid | - |
? | |
acetyl-CoA + 4-amino-3-hydroxybenzoic acid | - |
Mycobacterium tuberculosis | CoA + N-acetyl-4-amino-3-hydroxybenzoic acid | - |
? | |
acetyl-CoA + 4-chloroaniline | - |
Mycobacterium tuberculosis | CoA + N-acetyl-4-chloroaniline | - |
? | |
acetyl-CoA + 4-chloroaniline | - |
Mycobacterium tuberculosis | CoA + N-(4-chlorophenyl)acetamide | - |
? | |
acetyl-CoA + 4-fluoroaniline | - |
Mycobacterium tuberculosis | CoA + N-(4-fluorophenyl)acetamide | - |
? | |
acetyl-CoA + 4-fluoroaniline | - |
Mycobacterium tuberculosis | CoA + N-acetyl-4-fluoroaniline | - |
? | |
acetyl-CoA + 4-hydroxybenzhydrazide | - |
Mycobacterium tuberculosis | CoA + N'-acetyl-4-hydroxybenzohydrazide | - |
? | |
acetyl-CoA + 4-hydroxybenzhydrazide | - |
Mycobacterium tuberculosis | CoA + N-acetyl-4-hydroxybenzhydrazide | - |
? | |
acetyl-CoA + 4-methylaniline | - |
Mycobacterium tuberculosis | CoA + N-acetyl-4-methylaniline | - |
? | |
acetyl-CoA + 4-methylaniline | - |
Mycobacterium tuberculosis | CoA + N-(4-methylphenyl)acetamide | - |
? | |
acetyl-CoA + aniline | - |
Mycobacterium tuberculosis | CoA + N-acetyl-aniline | - |
? | |
acetyl-CoA + aniline | - |
Mycobacterium tuberculosis | CoA + N-phenylacetamide | - |
? | |
acetyl-CoA + anisidine | - |
Mycobacterium tuberculosis | CoA + N-acetyl-anisidine | - |
? | |
acetyl-CoA + anisidine | - |
Mycobacterium tuberculosis | CoA + N-(4-methoxyphenyl)acetamide | - |
? | |
acetyl-CoA + benzoic acid hydrazide | - |
Mycobacterium tuberculosis | CoA + N'-acetylbenzohydrazide | - |
? | |
acetyl-CoA + benzoic acid hydrazide | - |
Mycobacterium tuberculosis | CoA + N-acetyl-benzoic acid hydrazide | - |
? | |
acetyl-CoA + hydralazine | - |
Mycobacterium tuberculosis | CoA + N'-phthalazin-1-ylacetohydrazide | - |
? | |
acetyl-CoA + hydralazine | - |
Mycobacterium tuberculosis | CoA + N-acetyl-hydralazine | - |
? | |
acetyl-CoA + isoniazid | - |
Mycobacterium tuberculosis | CoA + N-acetyl-isoniazid | - |
? | |
acetyl-CoA + isoniazid | - |
Mycobacterium tuberculosis | CoA + N'-acetylpyridine-4-carbohydrazide | - |
? | |
acetyl-CoA + nicotinic acid hydrazide | - |
Mycobacterium tuberculosis | CoA + N'-acetylpyridine-3-carbohydrazide | - |
? | |
acetyl-CoA + nicotinic acid hydrazide | - |
Mycobacterium tuberculosis | CoA + N-acetyl-nicotinic acid hydrazide | - |
? | |
acetyl-CoA + p-aminobenzoic acid | - |
Mycobacterium tuberculosis | CoA + N-acetyl-p-aminobenzoic acid | - |
? | |
acetyl-CoA + p-aminobenzoic acid | - |
Mycobacterium tuberculosis | CoA + 4-(acetylamino)benzoic acid | - |
? | |
acetyl-CoA + sulfamethoxazole | - |
Mycobacterium tuberculosis | CoA + N-[4-[(5-methylisoxazol-3-yl)sulfamoyl]phenyl]acetamide | - |
? | |
additional information | results confirm that substituted anilines are excellent substrates and that TBNAT is ineffective in hydrazide N-acetylation | Mycobacterium tuberculosis | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
arylamine N-acetyltransferase | - |
Mycobacterium tuberculosis |
NAT | - |
Mycobacterium tuberculosis |
TBNAT | - |
Mycobacterium tuberculosis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Mycobacterium tuberculosis |
25 | - |
activity assay | Mycobacterium tuberculosis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.2 | - |
4-chloroaniline | - |
Mycobacterium tuberculosis | |
0.28 | - |
aniline | - |
Mycobacterium tuberculosis | |
0.6 | - |
nicotinic acid hydrazide | - |
Mycobacterium tuberculosis | |
0.7 | - |
p-Aminobenzoic acid | - |
Mycobacterium tuberculosis | |
0.8 | - |
4-methylaniline | - |
Mycobacterium tuberculosis | |
0.9 | - |
4-fluoroaniline | - |
Mycobacterium tuberculosis | |
1.9 | - |
benzoic acid hydrazide | - |
Mycobacterium tuberculosis | |
2.9 | - |
anisidine | - |
Mycobacterium tuberculosis | |
4.9 | - |
isoniazid | - |
Mycobacterium tuberculosis | |
10.9 | - |
4-hydroxybenzhydrazide | - |
Mycobacterium tuberculosis | |
21 | - |
2-amino-4-chlorophenol | - |
Mycobacterium tuberculosis | |
44 | - |
hydralazine | - |
Mycobacterium tuberculosis | |
51 | - |
2-Aminophenol | - |
Mycobacterium tuberculosis | |
66 | - |
4-amino-3-hydroxybenzoic acid | - |
Mycobacterium tuberculosis | |
70 | - |
2-amino-4-methylphenol | - |
Mycobacterium tuberculosis | |
94 | - |
3-Amino-4-hydroxybenzoic acid | - |
Mycobacterium tuberculosis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.7 | - |
assay at | Mycobacterium tuberculosis |
7.7 | - |
activity assay | Mycobacterium tuberculosis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
acetyl-CoA | - |
Mycobacterium tuberculosis |