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Literature summary for 2.3.3.13 extracted from

  • Huisman, F.H.; Squire, C.J.; Parker, E.J.
    Amino-acid substitutions at the domain interface affect substrate and allosteric inhibitor binding in alpha-isopropylmalate synthase from Mycobacterium tuberculosis (2013), Biochem. Biophys. Res. Commun., 433, 249-254.
    View publication on PubMed

Protein Variants

Protein Variants Comment Organism
D444A the mutant shows increased catalytic efficiency compared to the wild type enzyme Mycobacterium tuberculosis
D444R the mutant shows increased catalytic efficiency compared to the wild type enzyme Mycobacterium tuberculosis
D444Y the mutant shows decreased catalytic efficiency compared to the wild type enzyme Mycobacterium tuberculosis
R97A the mutant shows increased catalytic efficiency compared to the wild type enzyme Mycobacterium tuberculosis

Inhibitors

Inhibitors Comment Organism Structure
L-leucine non-competitive inhibition Mycobacterium tuberculosis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.005
-
3-methyl-2-oxobutanoate mutant enzyme D444A, at pH 7.5 and 25°C Mycobacterium tuberculosis
0.008
-
3-methyl-2-oxobutanoate mutant enzyme R97A, at pH 7.5 and 25°C Mycobacterium tuberculosis
0.009
-
3-methyl-2-oxobutanoate mutant enzyme D444R, at pH 7.5 and 25°C Mycobacterium tuberculosis
0.015
-
3-methyl-2-oxobutanoate wild type enzyme, at pH 7.5 and 25°C Mycobacterium tuberculosis
0.016
-
3-methyl-2-oxobutanoate mutant enzyme D444Y, at pH 7.5 and 25°C Mycobacterium tuberculosis
0.027
-
acetyl-CoA mutant enzyme D444R, at pH 7.5 and 25°C Mycobacterium tuberculosis
0.042
-
acetyl-CoA mutant enzyme D444A, at pH 7.5 and 25°C Mycobacterium tuberculosis
0.042
-
acetyl-CoA mutant enzyme D444Y, at pH 7.5 and 25°C Mycobacterium tuberculosis
0.14
-
acetyl-CoA wild type enzyme, at pH 7.5 and 25°C Mycobacterium tuberculosis
0.21
-
acetyl-CoA mutant enzyme R97A, at pH 7.5 and 25°C Mycobacterium tuberculosis

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ activates 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
-
?

Organism

Organism UniProt Comment Textmining
Mycobacterium tuberculosis P9WQB3
-
-

Purification (Commentary)

Purification (Comment) Organism
TALON affinity resin column chromatography 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
-
?

Subunits

Subunits Comment Organism
homodimer
-
Mycobacterium tuberculosis

Synonyms

Synonyms Comment Organism
alpha-IPMS
-
Mycobacterium tuberculosis
Alpha-isopropylmalate synthase
-
Mycobacterium tuberculosis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.1
-
3-methyl-2-oxobutanoate mutant enzyme D444A, at pH 7.5 and 25°C Mycobacterium tuberculosis
3.4
-
3-methyl-2-oxobutanoate mutant enzyme D444Y, at pH 7.5 and 25°C Mycobacterium tuberculosis
3.7
-
3-methyl-2-oxobutanoate wild type enzyme, at pH 7.5 and 25°C Mycobacterium tuberculosis
5.7
-
3-methyl-2-oxobutanoate mutant enzyme D444R, at pH 7.5 and 25°C Mycobacterium tuberculosis
7
-
3-methyl-2-oxobutanoate mutant enzyme R97A, at pH 7.5 and 25°C Mycobacterium tuberculosis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
27
-
acetyl-CoA wild type enzyme, at pH 7.5 and 25°C Mycobacterium tuberculosis
33
-
acetyl-CoA mutant enzyme R97A, at pH 7.5 and 25°C Mycobacterium tuberculosis
70
-
acetyl-CoA mutant enzyme D444A, at pH 7.5 and 25°C Mycobacterium tuberculosis
80
-
acetyl-CoA mutant enzyme D444Y, at pH 7.5 and 25°C Mycobacterium tuberculosis
210
-
acetyl-CoA mutant enzyme D444R, at pH 7.5 and 25°C Mycobacterium tuberculosis
210
-
3-methyl-2-oxobutanoate mutant enzyme D444Y, at pH 7.5 and 25°C Mycobacterium tuberculosis
250
-
3-methyl-2-oxobutanoate wild type enzyme, at pH 7.5 and 25°C Mycobacterium tuberculosis
600
-
3-methyl-2-oxobutanoate mutant enzyme D444A, at pH 7.5 and 25°C Mycobacterium tuberculosis
630
-
3-methyl-2-oxobutanoate mutant enzyme D444R, at pH 7.5 and 25°C Mycobacterium tuberculosis
900
-
3-methyl-2-oxobutanoate mutant enzyme R97A, at pH 7.5 and 25°C Mycobacterium tuberculosis