Application | Comment | Organism |
---|---|---|
medicine | alpha-isopropylmalate synthase is a target for the design of new antitubercular therapeutics | Mycobacterium tuberculosis |
Cloned (Comment) | Organism |
---|---|
into the vector pET28a for expression in Escherichia coli BL21DE3 pLysS cells | Mycobacterium tuberculosis |
Protein Variants | Comment | Organism |
---|---|---|
E218A | mutant, substitution at phylogenetically conserved active site residue near the interface of the catalytic and linker domains | Mycobacterium tuberculosis |
H379A | mutant, substitution at phylogenetically conserved active site residue near the interface of the catalytic and linker domains | Mycobacterium tuberculosis |
Y169F | mutant, substitution at phylogenetically conserved active site residue near the interface of the catalytic and linker domains | Mycobacterium tuberculosis |
Y410F | mutant, insensitive to L-leucine inhibition | Mycobacterium tuberculosis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
L-leucine | feedback inhibition | Mycobacterium tuberculosis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0005 | - |
alpha-ketoisovalerate | mutant Y410F | Mycobacterium tuberculosis | |
0.0011 | - |
acetyl-CoA | mutant Y410F | Mycobacterium tuberculosis | |
0.0016 | - |
alpha-ketoisovalerate | mutant E218A, enzyme displays substrate activation, unactivated form | Mycobacterium tuberculosis | |
0.0016 | - |
alpha-ketoisovalerate | mutant H379A, enzyme displays substrate activation, unactivated form | Mycobacterium tuberculosis | |
0.0023 | - |
acetyl-CoA | mutant H379A | Mycobacterium tuberculosis | |
0.0071 | - |
alpha-ketoisovalerate | wild-type | Mycobacterium tuberculosis | |
0.0095 | - |
acetyl-CoA | mutant E218A, enzyme displays substrate activation, unactivated form | Mycobacterium tuberculosis | |
0.0134 | - |
alpha-ketoisovalerate | mutant Y169F | Mycobacterium tuberculosis | |
0.019 | - |
alpha-ketoisovalerate | mutant H379A, enzyme displays substrate activation, activated form | Mycobacterium tuberculosis | |
0.024 | - |
acetyl-CoA | wild-type | Mycobacterium tuberculosis | |
0.039 | - |
alpha-ketoisovalerate | mutant E218A, enzyme displays substrate activation, activated form | Mycobacterium tuberculosis | |
0.04 | - |
acetyl-CoA | mutant Y169F | Mycobacterium tuberculosis | |
0.697 | - |
acetyl-CoA | mutant E218A, enzyme displays substrate activation, activated form | Mycobacterium tuberculosis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-CoA + 3-methyl-2-oxobutanoate + H2O | Mycobacterium tuberculosis | - |
(2S)-2-isopropylmalate + CoA | - |
? | |
acetyl-CoA + 3-methyl-2-oxobutanoate + H2O | Mycobacterium tuberculosis H37Rv | - |
(2S)-2-isopropylmalate + CoA | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycobacterium tuberculosis | P9WQB3 | - |
- |
Mycobacterium tuberculosis H37Rv | P9WQB3 | - |
- |
Purification (Comment) | Organism |
---|---|
on a nickel-NTA, a Mono-Q and a Phenyl Sepharose column | Mycobacterium tuberculosis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-CoA + 3-methyl-2-oxobutanoate + H2O | - |
Mycobacterium tuberculosis | (2S)-2-isopropylmalate + CoA | - |
? | |
acetyl-CoA + 3-methyl-2-oxobutanoate + H2O | - |
Mycobacterium tuberculosis H37Rv | (2S)-2-isopropylmalate + CoA | - |
? | |
alpha-ketoisovalerate + H2O + acetyl-CoA | - |
Mycobacterium tuberculosis | ? + CoA | - |
? | |
alpha-ketoisovalerate + H2O + acetyl-CoA | - |
Mycobacterium tuberculosis H37Rv | ? + CoA | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Alpha-isopropylmalate synthase | - |
Mycobacterium tuberculosis |
MtIPMS | - |
Mycobacterium tuberculosis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
activity assay | Mycobacterium tuberculosis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.022 | - |
alpha-ketoisovalerate | mutant E218A, enzyme displays substrate activation, activated form | Mycobacterium tuberculosis | |
0.037 | - |
alpha-ketoisovalerate | mutant E218A, enzyme displays substrate activation, unactivated form | Mycobacterium tuberculosis | |
0.058 | - |
acetyl-CoA | mutant E218A, enzyme displays substrate activation, unactivated form | Mycobacterium tuberculosis | |
0.075 | - |
alpha-ketoisovalerate | mutant Y410F | Mycobacterium tuberculosis | |
0.093 | - |
acetyl-CoA | mutant E218A, enzyme displays substrate activation, activated form | Mycobacterium tuberculosis | |
0.093 | - |
acetyl-CoA | mutant Y410F | Mycobacterium tuberculosis | |
0.7 | - |
alpha-ketoisovalerate | mutant H379A, enzyme displays substrate activation, unactivated form | Mycobacterium tuberculosis | |
1.483 | - |
alpha-ketoisovalerate | mutant H379A, enzyme displays substrate activation, activated form | Mycobacterium tuberculosis | |
2.53 | - |
alpha-ketoisovalerate | wild-type | Mycobacterium tuberculosis | |
2.55 | - |
acetyl-CoA | mutant H379A | Mycobacterium tuberculosis | |
2.717 | - |
acetyl-CoA | wild-type | Mycobacterium tuberculosis | |
4.75 | - |
alpha-ketoisovalerate | mutant Y169F | Mycobacterium tuberculosis | |
5.667 | - |
acetyl-CoA | mutant Y169F | Mycobacterium tuberculosis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
activity assay | Mycobacterium tuberculosis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
acetyl-CoA | - |
Mycobacterium tuberculosis |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.012 | - |
L-leucine | wild-type | Mycobacterium tuberculosis | |
0.018 | - |
L-leucine | mutant Y169F | Mycobacterium tuberculosis | |
0.019 | - |
L-leucine | mutant H379A | Mycobacterium tuberculosis | |
0.139 | - |
L-leucine | mutant E218A | Mycobacterium tuberculosis |