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Literature summary extracted from

  • Huisman, F.H.; Hunter, M.F.; Devenish, S.R.; Gerrard, J.A.; Parker, E.J.
    The C-terminal regulatory domain is required for catalysis by Neisseria meningitidis alpha-isopropylmalate synthase (2010), Biochem. Biophys. Res. Commun., 393, 168-173.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.3.13 into the vector pET-151 for expression in Escherichia coli BL21DE3 Star cells Neisseria meningitidis

Protein Variants

EC Number Protein Variants Comment Organism
2.3.3.13 E365Term mutant, truncated form of NmeIPMS, loss of the regulatory domain results in a loss of the ability to catalyse the adol reaction, although the enzyme is still able to slowly hydrolyse AcCoA Neisseria meningitidis

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.3.3.13 (S)-alpha-hydroxyisovalerate
-
 Neisseria meningitidis
2.3.3.13 3,3-dibromopyruvate
-
 Neisseria meningitidis
2.3.3.13 3-Bromopyruvate
-
 Neisseria meningitidis
2.3.3.13 leucine
-
 Neisseria meningitidis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.3.13 0.027
-
 3-methyl-2-oxopentanoate pH 7.5, 25°C Neisseria meningitidis
2.3.3.13 0.3
-
 3-Fluoropyruvate pH 7.5, 25°C Neisseria meningitidis
2.3.3.13 0.63
-
 pyruvate pH 7.5, 25°C Neisseria meningitidis
2.3.3.13 3.3
-
 alpha-Ketobutyrate pH 7.5, 25°C Neisseria meningitidis
2.3.3.13 5.4
-
 alpha-ketoisovalerate pH 7.5, 25°C Neisseria meningitidis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.3.3.13 Cd2+
-
 Neisseria meningitidis
2.3.3.13 Zn2+
-
 Neisseria meningitidis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.3.3.13 39930
-
 E365Term mutant, theoretical Neisseria meningitidis
2.3.3.13 40000
-
 E365Term mutant, determined by SDS-PAGE Neisseria meningitidis
2.3.3.13 56000
-
 wild-type enzyme, determined by SDS-PAGE Neisseria meningitidis
2.3.3.13 56030
-
 wild-type enzyme, theoretical Neisseria meningitidis

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 Neisseria meningitidis
-
 (2S)-2-isopropylmalate + CoA
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.3.3.13 Neisseria meningitidis Q9JZG1
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.3.13 using Talon Superflow Metal Affinity resin, the His-tag is removed by TEV protease, anion exchange chromatography on a SourceQ column is applied Neisseria meningitidis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.3.13 2-oxobutyrate + acetyl-CoA + H2O
-
 Neisseria meningitidis ?
-
 ?
2.3.3.13 3-fluoropyruvate + acetyl-CoA + H2O
-
 Neisseria meningitidis ?
-
 ?
2.3.3.13 3-methyl-2-oxopentanoate + acetyl-CoA + H2O
-
 Neisseria meningitidis ?
-
 ?
2.3.3.13 acetyl-CoA + 3-methyl-2-oxobutanoate + H2O
-
 Neisseria meningitidis (2S)-2-isopropylmalate + CoA
-
 ?
2.3.3.13 alpha-ketoisovalerate + acetyl-CoA + H2O
-
 Neisseria meningitidis alpha-isopropylmalate + CoA
-
 ?
2.3.3.13 pyruvate + acetyl-CoA + H2O
-
 Neisseria meningitidis ?
-
 ?

Subunits

EC Number Subunits Comment Organism
2.3.3.13 homodimer
-
 Neisseria meningitidis

Synonyms

EC Number Synonyms Comment Organism
2.3.3.13 alpha-IPMS
-
 Neisseria meningitidis
2.3.3.13 Alpha-isopropylmalate synthase
-
 Neisseria meningitidis
2.3.3.13 NmeIPMS
-
 Neisseria meningitidis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.3.3.13 25
-
 activity assay Neisseria meningitidis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.3.3.13 0.035
-
 alpha-ketoisovalerate pH 7.5, 25°C Neisseria meningitidis
2.3.3.13 0.28
-
 3-methyl-2-oxopentanoate pH 7.5, 25°C Neisseria meningitidis
2.3.3.13 1.36
-
 alpha-Ketobutyrate pH 7.5, 25°C Neisseria meningitidis
2.3.3.13 1.5
-
 pyruvate pH 7.5, 25°C Neisseria meningitidis
2.3.3.13 105
-
 3-Fluoropyruvate pH 7.5, 25°C Neisseria meningitidis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.3.13 7.5
-
 activity assay Neisseria meningitidis

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.3.3.13 0.03
-
 3,3-dibromopyruvate pH 7.5, 25°C Neisseria meningitidis
2.3.3.13 0.06
-
 3-Bromopyruvate pH 7.5, 25°C Neisseria meningitidis
2.3.3.13 2.5
-
 (S)-alpha-hydroxyisovalerate pH 7.5, 25°C Neisseria meningitidis