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Literature summary for 2.3.3.13 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)

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

Protein Variants

Protein Variants Comment Organism
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

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

KM Value [mM]

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

Metals/Ions

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

Molecular Weight [Da]

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

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
acetyl-CoA + 3-methyl-2-oxobutanoate + H2O Neisseria meningitidis
-
(2S)-2-isopropylmalate + CoA
-
?

Organism

Organism UniProt Comment Textmining
Neisseria meningitidis Q9JZG1
-
-

Purification (Commentary)

Purification (Comment) Organism
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)

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

Subunits

Subunits Comment Organism
homodimer
-
Neisseria meningitidis

Synonyms

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

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

pH Optimum

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

Ki Value [mM]

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