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Literature summary 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)

EC Number Cloned (Comment) Organism
2.3.3.13 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
2.3.3.13 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)

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

Protein Variants

EC Number Protein Variants Comment Organism
2.3.3.13 additional information the E365Term protein lacks the C-terminal allosteric regulatory domain of the wild type enzyme and is inactive Neisseria meningitidis
2.3.3.13 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.3.13 0.012
-
2-oxo-3-methylbutanoate apparent value, full-length enzyme, in 50 mM HEPES (pH 7.5), at 25°C Mycobacterium tuberculosis
2.3.3.13 0.03
-
2-oxo-3-methylbutanoate apparent value, full-length enzyme, in 50 mM HEPES (pH 7.5), at 25°C Neisseria meningitidis
2.3.3.13 0.035
-
acetyl-CoA apparent value, full-length enzyme, in 50 mM HEPES (pH 7.5), at 25°C Neisseria meningitidis
2.3.3.13 0.136
-
acetyl-CoA apparent value, full-length enzyme, in 50 mM HEPES (pH 7.5), at 25°C Mycobacterium tuberculosis

Metals/Ions

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

Organism

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

Purification (Commentary)

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

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.3.13 2-oxo-3-methylbutanoate + acetyl-CoA + H2O
-
Mycobacterium tuberculosis 3-hydroxy-4-methyl-3-carboxypentanoate + CoA
-
ir
2.3.3.13 2-oxo-3-methylbutanoate + acetyl-CoA + H2O
-
Neisseria meningitidis 3-hydroxy-4-methyl-3-carboxypentanoate + CoA
-
ir
2.3.3.13 2-oxo-3-methylbutanoate + acetyl-CoA + H2O
-
Mycobacterium tuberculosis H37Rv 3-hydroxy-4-methyl-3-carboxypentanoate + CoA
-
ir

Subunits

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

Synonyms

EC Number Synonyms Comment Organism
2.3.3.13 alpha-IPMS
-
Mycobacterium tuberculosis
2.3.3.13 alpha-IPMS
-
Neisseria meningitidis
2.3.3.13 alpha-isopropylmalate synthase I
-
Mycobacterium tuberculosis
2.3.3.13 alpha-isopropylmalate synthase I
-
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 3.5
-
acetyl-CoA full-length enzyme, in 50 mM HEPES (pH 7.5), at 25°C Mycobacterium tuberculosis
2.3.3.13 3.5
-
2-oxo-3-methylbutanoate full-length enzyme, in 50 mM HEPES (pH 7.5), at 25°C Mycobacterium tuberculosis
2.3.3.13 13
-
acetyl-CoA full-length enzyme, in 50 mM HEPES (pH 7.5), at 25°C Neisseria meningitidis
2.3.3.13 13
-
2-oxo-3-methylbutanoate full-length enzyme, in 50 mM HEPES (pH 7.5), at 25°C Neisseria meningitidis