3-Hydroxy-3-methylglutaryldithio-CoA: utility of an alternative substrate in elucidation of a role for HMG-CoA lyase's cation activator

Paul W. Hruz, Vernon E. Anderson, Henry M. Miziorko

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11 Scopus citations


(S)-(3-Hydroxy-3-methyl-1-thionoglutaryl)-Coenzyme A (HMG[ = S]CoA), a dithioester analog of (S)-(3-hydroxy-3-methylglutaryl)-CoA (HMG-CoA), acts as an efficient alternative substrate for avian HMG-CoA lyase. Detection of product formation by HPLC, UV absorbance and coupled enzyme assays indicates that HMG[ = S]CoA cleavage yields acetyl[ = S]CoA and acetoacetate. HMG[ = S]CoA binds to the lyase with a Km of 13 μM and undergoes the cleavage reaction at a maximal rate which is 20% of that observed with HMG-CoA. The enzyme-catalyzed cleavage of both HMG-CoA and HMG[ = S]CoA is stimulated by the divalent cations Mg2+ and Mn2+. Mg2+ produces a 2-fold higher stimulation of HMG-CoA cleavage than that observed with Mn2+. In contrast, stimulation of HMG[ = S]CoA cleavage is nearly seven times higher with Mn2+ than with Mg2+. Not only is the stimulation of enzymatic activity dependent on the cation, but also the Km values for Mg2+ and Mn2+ are dependent upon the substrate used. In contrast, the Km values for HMG-CoA and HMG[ = S]CoA are not markedly dependent on the identity of the divalent cation. These results are compatible with the initial formation of a binary enzyme-substrate complex prior to binding of the divalent cation to produce a catalytically active enzyme-substrate-metal ternary complex.

Original languageEnglish
Pages (from-to)149-154
Number of pages6
JournalBiochimica et Biophysica Acta (BBA)/Protein Structure and Molecular
Issue number1-2
StatePublished - Mar 5 1993


  • Divalent cation
  • HMG-CoA lyase
  • HMG[ = S]CoA


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