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.
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 |