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Literature summary extracted from

  • Fu, Z.; Runquist, J.A.; Forouhar, F.; Hussain, M.; Hunt, J.F.; Miziorko, H.M.; Kim, J.J.
    Crystal structure of human HMG-CoA lyase: Insights into catalysis and the molecular basis for hydroxymethylglutaric aciduria (2005), J. Biol. Chem., 281, 7526-7532.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.1.3.4 expressed in Escherichia coli Homo sapiens

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
4.1.3.4 crystal structure at 2.1 A resolution of the recombinant mitochondrial HMG-CoA lyase containing a bound activator cation and 3-hydroxyglutarate. For crystallization experiments, the enzyme is diluted to 5 –6 mg/ml. The competitive inhibitor hydroxyglutaryl-CoA is added to the diluted protein at about 1mM concentration. The inhibitor is necessary for generation of uniform diffraction quality crystals. Crystals sufficient for X-ray studies are obtained using an equilibration buffer of 0.1 M Hepes, pH 7.5, 60 mM MgCl2,and 15% polyethylene glycol 8K. The enzyme is mixed 1:1 with the equilibration buffer using sitting drop trays at 19°C. Crystals belong to the monoclinic space group C2 with unit cell parameters a = 197.0 A, b = 117.1 A, c = 86.8 A, and beta = 112.5°. Six monomers are found in the asymmetric unit Homo sapiens
4.1.3.4 sitting drop vapour diffusion method with 0.1 M HEPES, pH 7.5, 60 mM MgCl2, and 15% polyethylene glycol 8K Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
4.1.3.4 D204N decreased activity Homo sapiens
4.1.3.4 D42A exhibits catalytic rates diminished by 130000fold Homo sapiens
4.1.3.4 D42H decreased activity Homo sapiens
4.1.3.4 E279K unstable recombinant protein Homo sapiens
4.1.3.4 H233A exhibits catalytic rates diminished by 6400fold Homo sapiens
4.1.3.4 H233R decreased activity Homo sapiens
4.1.3.4 R41Q decreased activity Homo sapiens
4.1.3.4 S201Y no activity Homo sapiens
4.1.3.4 S75R no activity Homo sapiens

Inhibitors

EC Number Inhibitors Comment Organism Structure
4.1.3.4 3-hydroxyglutaryl-CoA competitive; competitive inhibitor Homo sapiens

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
4.1.3.4 mitochondrion
-
Homo sapiens 5739
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
4.1.3.4 additional information identification of His233, His235, ASp42, and Asn275 as metal-binding ligands Homo sapiens

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
4.1.3.4 32000
-
SDS-PAGE Homo sapiens

Organism

EC Number Organism UniProt Comment Textmining
4.1.3.4 Homo sapiens
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.1.3.4 Q-Sepharose column chromatography, phenyl-agarose column chromatography, and Superose 12 column chromatography Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.1.3.4 (S)-3-Hydroxy-3-methylglutaryl-CoA
-
Homo sapiens Acetyl-CoA + acetoacetate
-
?

Subunits

EC Number Subunits Comment Organism
4.1.3.4 hexamer X-ray crystallography, the hexamer consists of three dimers which are supposed to be the physiological enzyme form in solution Homo sapiens

Synonyms

EC Number Synonyms Comment Organism
4.1.3.4 3-hydroxy-3-methylglutarate-CoA lyase
-
Homo sapiens
4.1.3.4 HMG-CoA lyase
-
Homo sapiens