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Literature summary 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)

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

Protein Variants

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

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.3.3.13 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
2.3.3.13 additional information not inhibited by EDTA Mycobacterium tuberculosis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.3.13 0.097
-
 acetyl-CoA mutant enzyme D81A, at pH 7.5 and 25°C Mycobacterium tuberculosis
2.3.3.13 0.123
-
 acetyl-CoA mutant enzyme N321A, at pH 7.5 and 25°C Mycobacterium tuberculosis
2.3.3.13 0.163
-
 acetyl-CoA mutant enzyme D81E/N321I, at pH 7.5 and 25°C Mycobacterium tuberculosis
2.3.3.13 0.233
-
 acetyl-CoA mutant enzyme D81E, at pH 7.5 and 25°C Mycobacterium tuberculosis
2.3.3.13 0.253
-
 acetyl-CoA wild type enzyme, at pH 7.5 and 25°C Mycobacterium tuberculosis
2.3.3.13 0.262
-
 acetyl-CoA mutant enzyme N321I, at pH 7.5 and 25°C Mycobacterium tuberculosis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.3.3.13 72000
-
 2 * 72000, SDS-PAGE Mycobacterium tuberculosis
2.3.3.13 171000
-
 gel filtration Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.3.3.13 acetyl-CoA + 3-methyl-2-oxobutanoate + H2O Mycobacterium tuberculosis
-
 (2S)-2-isopropylmalate + CoA
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.3.3.13 Mycobacterium tuberculosis
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.3.13 HisTrap column chromatography Mycobacterium tuberculosis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.3.13 acetyl-CoA + 3-methyl-2-oxobutanoate + H2O
-
 Mycobacterium tuberculosis (2S)-2-isopropylmalate + CoA
-
 ?

Subunits

EC Number Subunits Comment Organism
2.3.3.13 homodimer 2 * 72000, SDS-PAGE Mycobacterium tuberculosis

Synonyms

EC Number Synonyms Comment Organism
2.3.3.13 alpha-IPMS
-
 Mycobacterium tuberculosis
2.3.3.13 Alpha-isopropylmalate synthase
-
 Mycobacterium tuberculosis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.3.3.13 0.05
-
 acetyl-CoA mutant enzyme D81A, at pH 7.5 and 25°C Mycobacterium tuberculosis
2.3.3.13 0.12
-
 acetyl-CoA mutant enzyme D81E, at pH 7.5 and 25°C Mycobacterium tuberculosis
2.3.3.13 0.13
-
 acetyl-CoA mutant enzyme D81E/N321I, at pH 7.5 and 25°C Mycobacterium tuberculosis
2.3.3.13 0.15
-
 acetyl-CoA mutant enzyme N321A, at pH 7.5 and 25°C Mycobacterium tuberculosis
2.3.3.13 0.15
-
 acetyl-CoA mutant enzyme N321I, at pH 7.5 and 25°C Mycobacterium tuberculosis
2.3.3.13 0.32
-
 acetyl-CoA wild type enzyme, at pH 7.5 and 25°C Mycobacterium tuberculosis