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Literature summary for 2.3.3.13 extracted from

  • Frantom, P.A.; Birman, Y.; Hays, B.N.; Casey, A.K.
    An evolutionarily conserved alternate metal ligand is important for activity in alpha-isopropylmalate synthase from Mycobacterium tuberculosis (2014), Biochim. Biophys. Acta, 1844, 1784-1789.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3)pLysS cells Mycobacterium tuberculosis

Protein Variants

Protein Variants Comment Organism
D81A the mutant shows strongly decreased activity compared to the wild type enzyme Mycobacterium tuberculosis
D81E the mutant shows strongly decreased activity compared to the wild type enzyme Mycobacterium tuberculosis
D81E/N321I the mutant shows strongly decreased activity compared to the wild type enzyme Mycobacterium tuberculosis
N321A the mutant shows strongly decreased activity compared to the wild type enzyme Mycobacterium tuberculosis
N321I the mutant shows strongly decreased activity compared to the wild type enzyme Mycobacterium tuberculosis

Inhibitors

Inhibitors Comment Organism Structure
Mg2+ hydrolysis activity of the wild type enzyme is inhibited by increasing concentrations of Mg2+ with IC50 values in the low millimolar range Mycobacterium tuberculosis
additional information not inhibited by EDTA Mycobacterium tuberculosis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.097
-
acetyl-CoA mutant enzyme D81A, at pH 7.5 and 25°C Mycobacterium tuberculosis
0.123
-
acetyl-CoA mutant enzyme N321A, at pH 7.5 and 25°C Mycobacterium tuberculosis
0.163
-
acetyl-CoA mutant enzyme D81E/N321I, at pH 7.5 and 25°C Mycobacterium tuberculosis
0.233
-
acetyl-CoA mutant enzyme D81E, at pH 7.5 and 25°C Mycobacterium tuberculosis
0.253
-
acetyl-CoA wild type enzyme, at pH 7.5 and 25°C Mycobacterium tuberculosis
0.262
-
acetyl-CoA mutant enzyme N321I, at pH 7.5 and 25°C Mycobacterium tuberculosis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
72000
-
2 * 72000, SDS-PAGE Mycobacterium tuberculosis
171000
-
gel filtration 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
-
-
-

Purification (Commentary)

Purification (Comment) Organism
HisTrap 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 2 * 72000, SDS-PAGE 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
0.05
-
acetyl-CoA mutant enzyme D81A, at pH 7.5 and 25°C Mycobacterium tuberculosis
0.12
-
acetyl-CoA mutant enzyme D81E, at pH 7.5 and 25°C Mycobacterium tuberculosis
0.13
-
acetyl-CoA mutant enzyme D81E/N321I, at pH 7.5 and 25°C Mycobacterium tuberculosis
0.15
-
acetyl-CoA mutant enzyme N321A, at pH 7.5 and 25°C Mycobacterium tuberculosis
0.15
-
acetyl-CoA mutant enzyme N321I, at pH 7.5 and 25°C Mycobacterium tuberculosis
0.32
-
acetyl-CoA wild type enzyme, at pH 7.5 and 25°C Mycobacterium tuberculosis