Inactivation of D-3-hydroxybutyrate dehydrogenase by fumaroyl bis(methyl phosphate).
Fumaroyl bis(methyl phosphate) reacts with the NADH-dependent enzyme, D-3-hydroxybutyrate dehydrogenase, leading to irreversible inactivation. The bifunctional reagent cross-links the subunits of the enzyme. The inactivation is subject to saturation and protection by substrate, consistent with the reaction occurring at the active site. The stoichiometry of inactivation indicates two active sites undergo reaction with each equivalent of reagent. These results indicate that the dimeric enzyme has contiguous active sites. The reagent is likely to react with an active site lysine, consistent with previous suggestions.
Kluger
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R
Lash Miller Laboratories, Department of Chemistry, University of Toronto, Canada.
Bearne
S L
SL
D001665:Binding Sites; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005650:Fumarates / Q000494:pharmacology*; D006884:Hydroxybutyrate Dehydrogenase / Q000037:antagonists & inhibitors* / Q000737:chemistry; D007202:Indicators and Reagents; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D010755:Organophosphates / Q000494:pharmacology*
D005650:Fumarates; D007202:Indicators and Reagents; D046911:Macromolecular Substances; D010755:Organophosphates; C091057:fumaroyl bis(methylphosphate); D006884:Hydroxybutyrate Dehydrogenase