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Literature summary for 2.3.3.13 extracted from

  • Huisman, F.H.; Koon, N.; Bulloch, E.M.; Baker, H.M.; Baker, E.N.; Squire, C.J.; Parker, E.J.
    Removal of the C-terminal regulatory domain of alpha-isopropylmalate synthase disrupts functional substrate binding (2012), Biochemistry, 51, 2289-2297.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
truncated protein E365Term is expressed in Escherichia coli BL21(DE3) Star cells and in Escherichia coli strain BW25113 lacking the alpha-isopropylmalate synthase gene Neisseria meningitidis
truncated protein LeuA425 is expressed in Escherichia coli Rosetta 2 cells and in Escherichia coli strain BW25113 lacking the alpha-isopropylmalate synthase gene Mycobacterium tuberculosis

Crystallization (Commentary)

Crystallization (Comment) Organism
truncated protein E365Term, sitting drop vapor diffusion method, using 0.2 M magnesium acetate and 18% (w/v) mPEG 3350 Neisseria meningitidis
truncated protein LeuA425, sitting drop vapor diffusion method, using 24% (w/v) PEG 3350 and 275 mM lithium nitrate Mycobacterium tuberculosis

Protein Variants

Protein Variants Comment Organism
additional information the E365Term protein lacks the C-terminal allosteric regulatory domain of the wild type enzyme and is inactive Neisseria meningitidis
additional information the LeuA425 protein lacks the C-terminal allosteric regulatory domain of the wild type enzyme and is inactive Mycobacterium tuberculosis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.012
-
2-oxo-3-methylbutanoate apparent value, full-length enzyme, in 50 mM HEPES (pH 7.5), at 25°C Mycobacterium tuberculosis
0.03
-
2-oxo-3-methylbutanoate apparent value, full-length enzyme, in 50 mM HEPES (pH 7.5), at 25°C Neisseria meningitidis
0.035
-
acetyl-CoA apparent value, full-length enzyme, in 50 mM HEPES (pH 7.5), at 25°C Neisseria meningitidis
0.136
-
acetyl-CoA apparent value, full-length enzyme, in 50 mM HEPES (pH 7.5), at 25°C Mycobacterium tuberculosis

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+ contains two Mn2+ ion Mycobacterium tuberculosis
Mn2+ contains two Mn2+ ions Neisseria meningitidis
Zn2+ contains one Zn2+ ion Mycobacterium tuberculosis
Zn2+ contains one Zn2+ ion Neisseria meningitidis

Organism

Organism UniProt Comment Textmining
Mycobacterium tuberculosis P9WQB3
-
-
Mycobacterium tuberculosis H37Rv P9WQB3
-
-
Neisseria meningitidis Q9JZG1
-
-

Purification (Commentary)

Purification (Comment) Organism
Talon metal affinity resin column chromatography and Sephacryl 200 gel filtration Mycobacterium tuberculosis
Talon metal affinity resin column chromatography and Sephacryl 200 gel filtration Neisseria meningitidis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-oxo-3-methylbutanoate + acetyl-CoA + H2O
-
Mycobacterium tuberculosis 3-hydroxy-4-methyl-3-carboxypentanoate + CoA
-
ir
2-oxo-3-methylbutanoate + acetyl-CoA + H2O
-
Neisseria meningitidis 3-hydroxy-4-methyl-3-carboxypentanoate + CoA
-
ir
2-oxo-3-methylbutanoate + acetyl-CoA + H2O
-
Mycobacterium tuberculosis H37Rv 3-hydroxy-4-methyl-3-carboxypentanoate + CoA
-
ir

Subunits

Subunits Comment Organism
homodimer x-ray crystallography Mycobacterium tuberculosis
homodimer x-ray crystallography Neisseria meningitidis

Synonyms

Synonyms Comment Organism
alpha-IPMS
-
Mycobacterium tuberculosis
alpha-IPMS
-
Neisseria meningitidis
alpha-isopropylmalate synthase I
-
Mycobacterium tuberculosis
alpha-isopropylmalate synthase I
-
Neisseria meningitidis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5
-
acetyl-CoA full-length enzyme, in 50 mM HEPES (pH 7.5), at 25°C Mycobacterium tuberculosis
3.5
-
2-oxo-3-methylbutanoate full-length enzyme, in 50 mM HEPES (pH 7.5), at 25°C Mycobacterium tuberculosis
13
-
acetyl-CoA full-length enzyme, in 50 mM HEPES (pH 7.5), at 25°C Neisseria meningitidis
13
-
2-oxo-3-methylbutanoate full-length enzyme, in 50 mM HEPES (pH 7.5), at 25°C Neisseria meningitidis