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Literature summary for 2.3.3.13 extracted from

  • de Carvalho, L.P.; Frantom, P.A.; Argyrou, A.; Blanchard, J.S.
    Kinetic evidence for interdomain communication in the allosteric regulation of alpha-isopropylmalate synthase from Mycobacterium tuberculosis (2009), Biochemistry, 48, 1996-2004.
    View publication on PubMedView publication on EuropePMC

Application

Application Comment Organism
medicine alpha-isopropylmalate synthase is a target for the design of new antitubercular therapeutics Mycobacterium tuberculosis

Cloned(Commentary)

Cloned (Comment) Organism
into the vector pET28a for expression in Escherichia coli BL21DE3 pLysS cells Mycobacterium tuberculosis

Protein Variants

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

Inhibitors Comment Organism Structure
L-leucine feedback inhibition Mycobacterium tuberculosis

KM Value [mM]

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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Mycobacterium tuberculosis P9WQB3
-
-
Mycobacterium tuberculosis H37Rv P9WQB3
-
-

Purification (Commentary)

Purification (Comment) Organism
on a nickel-NTA, a Mono-Q and a Phenyl Sepharose column Mycobacterium tuberculosis

Substrates and Products (Substrate)

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

Synonyms Comment Organism
Alpha-isopropylmalate synthase
-
Mycobacterium tuberculosis
MtIPMS
-
Mycobacterium tuberculosis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
activity assay Mycobacterium tuberculosis

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
activity assay Mycobacterium tuberculosis

Cofactor

Cofactor Comment Organism Structure
acetyl-CoA
-
Mycobacterium tuberculosis

Ki Value [mM]

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