|Gene:||mvaB||Accession Number: G-9191 (MetaCyc)|
Species: Pseudomonas mevalonii
Subunit composition of
hydroxymethylglutaryl-CoA lyase = [MvaB]2
hydroxymethylglutaryl-CoA lyase monomer = MvaB
The activity of hydroxymethylglutaryl-CoA lyase monomer has been initially documented in 1955 by Bachhawat et al from mammalian tissues, including pig heart, liver and kidney, and pigeon liver. They partially purified the enzyme from pig heart [Bachhawat55]. In 1967 the activity was first demonstrated in bacteria by Siddiqi and Rodwell, who isolated a (R)-mevalonate degrading bacterium from soil [Siddiqi67]. The organism, known only as Actinomycete S4, was capable of growth using (R)-mevalonate as the sole carbon source, converting it to acetoacetate and acetyl-CoA. Siddiqi and Rodwell were able to show that (S)-3-hydroxy-3-methylglutaryl-CoA was the intermediate in the process.
The enzyme was purified 650-fold from cell extracts, to a specific activity of 22 μmol acetyl-CoA formed per min per mg protei. The purified enzyme, which was monomeric, required either Mn2+ or Mg2+ for activityn [Scher89].
The gene encoding the enzyme was discovered in 1989 by Anderson and Rodwell, who were working with a different (R)-mevalonate degrading bacterium, Pseudomonas mevalonii [Anderson89]. The gene, which resides in an operon with the gene encoding hydroxymethylglutaryl-CoA reductase, was cloned and overexpressed in Escherichia coli.
In a subsequent study, atomic absorption and EPR analyses of isolated enzyme indicated the presence of tightly bound copper. Copper-enriched enzyme displayed enhanced thermal stability [Narasimhan92]. Study of the copper binding site by multifrequency electron spin resonance spectroscopy suggested that at least two (and probably only two) nitrogen donor atoms are liganded to the tightly bound copper, and that the enzyme is composed of two identical subunits [Narasimhan94].
Gene Citations: [Beach89]
Molecular Weight of Polypeptide: 31.609 kD (from nucleotide sequence), 31.6 kD (experimental) [Anderson89 ]
Relationship Links: Entrez-Nucleotide:PART-OF:M24016 , InterPro:IN-FAMILY:IPR000138 , InterPro:IN-FAMILY:IPR000891 , InterPro:IN-FAMILY:IPR013785 , InterPro:IN-FAMILY:IPR027167 , Panther:IN-FAMILY:PTHR10277:SF1 , Pfam:IN-FAMILY:PF00682 , Prosite:IN-FAMILY:PS01062 , Prosite:IN-FAMILY:PS50991
|Cellular Component:||GO:0005829 - cytosol [Anderson89]|
Enzymatic reaction of: hydroxymethylglutaryl-CoA lyase
EC Number: 188.8.131.52
The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the Enzyme Commission system.
The reaction is favored in the direction shown.
pH(opt): 8.8 [Scher89]
Anderson89: Anderson DH, Rodwell VW (1989). "Nucleotide sequence and expression in Escherichia coli of the 3-hydroxy-3-methylglutaryl coenzyme A lyase gene of Pseudomonas mevalonii." J Bacteriol 171(12);6468-72. PMID: 2687236
Bachhawat55: Bachhawat, B.K., Robinson, W.G., Coon, M.J. (1955). "The enzymatic cleavage of beta-hydroxy-beta-methylglutaryl coenzyme A to acetoacetate and acetyl coenzyme A." J Biol Chem 216(2);727-36. PMID: 13271348
Beach89: Beach MJ, Rodwell VW (1989). "Cloning, sequencing, and overexpression of mvaA, which encodes Pseudomonas mevalonii 3-hydroxy-3-methylglutaryl coenzyme A reductase." J Bacteriol 171(6);2994-3001. PMID: 2656635
Hruz92: Hruz PW, Narasimhan C, Miziorko HM (1992). "3-Hydroxy-3-methylglutaryl coenzyme A lyase: affinity labeling of the Pseudomonas mevalonii enzyme and assignment of cysteine-237 to the active site." Biochemistry 31(29);6842-7. PMID: 1637819
Narasimhan92: Narasimhan C, Miziorko HM (1992). "Pseudomonas mevalonii 3-hydroxy-3-methylglutaryl-CoA lyase: characterization of the isolated recombinant protein and investigation of the enzyme's cation requirements." Biochemistry 31(45);11224-30. PMID: 1332752
Narasimhan94: Narasimhan C, Antholine WE, Miziorko HM (1994). "P. mevalonii 3-hydroxy-3-methylglutaryl-CoA lyase: electron paramagnetic resonance investigation of the copper binding site." Arch Biochem Biophys 312(2);467-73. PMID: 7913596
Narasimhan95: Narasimhan C, Roberts JR, Miziorko HM (1995). "Pseudomonas mevalonii 3-hydroxy-3-methylglutaryl-CoA lyase: testing the function of the active site cysteine by site-directed mutagenesis." Biochemistry 34(31);9930-5. PMID: 7632692
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