(E)-3-Cyanophosphoenolpyruvate, a new inhibitor of phosphoenolpyruvate-dependent enzymes.
(E)-3-Cyanophosphoenolpyruvate has been synthesized by reacting dimethyl chlorophosphate with the potassium enolate of ethyl cyanopyruvate. The resulting trialkyl ester was deesterified with bromotrimethylsilane followed by potassium hydroxide. Subsequent treatment with Dowex-50-H+ resin and cyclohexylamine afforded the tricyclohexylammonium salt; only the E geometric isomer was obtained. This compound can be photoisomerized to a 70:30 E:Z mixture. (E)-3-Cyanophosphoenolpyruvate is an excellent competitive inhibitor of phosphoenolpyruvate carboxylase [KI(Mn2+) = 16 microM, KI(Mg2+) = 1360 microM], pyruvate kinase [KI(Mn2+) = 0.085 microM, KI(Mg2+) = 0.76 microM], and enolase [KI(Mn2+) = 360 microM, KI(Mg2+) = 280 microM]. The compound is a substrate for pyruvate kinase (Vmax approximately 1% of phosphoenolpyruvate rate), but not for the other two enzymes. No irreversible inactivation is observed with phosphoenolpyruvate carboxylase of pyruvate kinase.
D000818:Animals; D001667:Binding, Competitive; D004791:Enzyme Inhibitors / Q000138:chemical synthesis*; D066298:In Vitro Techniques; D007700:Kinetics; D010728:Phosphoenolpyruvate / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000494:pharmacology; D010730:Phosphoenolpyruvate Carboxylase / Q000037:antagonists & inhibitors; D010751:Phosphopyruvate Hydratase / Q000037:antagonists & inhibitors; D011770:Pyruvate Kinase / Q000037:antagonists & inhibitors / Q000378:metabolism; D011817:Rabbits; D013379:Substrate Specificity
D004791:Enzyme Inhibitors; D010728:Phosphoenolpyruvate; C048060:3-cyanophosphoenolpyruvate; D011770:Pyruvate Kinase; D010730:Phosphoenolpyruvate Carboxylase; D010751:Phosphopyruvate Hydratase