Phosphoenolpyruvate carboxylase of Escherichia coli. Specificity of some compounds as activators at the site for fructose 1,6-bisphosphate, one of the allosteric effectors.
An investigation was performed to elucidate some unusual phenomena which had been observed with phosphoenolpyruvate (PEP) carboxylase [EC 4.1.1.31] of Escherichia coli. (i) Fructose 1,6-bisphosphate (Fru-1,6-P2) and GTP--the allosteric activators--were competitive with each other in the activation. (ii) Some analogs of PEP such as DL-2-phospholactate and 2-phosphoglycolate, which behaved as inhibitors in the presence of the activator (acetyl-CoA or dioxane), activated the enzyme to some extent in the absence of the activator. (iii) Ammonium sulfate deprived the enzyme of sensitivity to Fru-1,6-P2 or GTP but had no effect on the sensitivity to other effectors. It was found that the activation by the analogs was lost upon desensitization of the enzyme to Fru-1,6-P2 by reaction with 2,4,6-trinitrobenzene sulfonate. The activation by the analogs was not observed in the presence of 200 mM ammonium sulfate. In the presence of lower concentrations (0.1 mM) of PEP, ammonium sulfate activated the enzyme at concentrations less than 700 mM but had an inhibitory effect on the desensitized enzyme. These findings suggest that the unusual phenomena described above are a result of binding of the phosphate esters and sulfate ions with the Fru-1,6-P2 site of the enzyme or the active site depending on the reaction conditions.
D000495:Allosteric Site; D000645:Ammonium Sulfate / Q000494:pharmacology; D002262:Carboxy-Lyases / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D004926:Escherichia coli / Q000201:enzymology*; D005635:Fructosediphosphates / Q000494:pharmacology*; D006598:Hexosediphosphates / Q000494:pharmacology*; D010728:Phosphoenolpyruvate / Q000031:analogs & derivatives / Q000494:pharmacology; D010730:Phosphoenolpyruvate Carboxylase / Q000378:metabolism*; D013237:Stereoisomerism; D014302:Trinitrobenzenesulfonic Acid / Q000494:pharmacology
D005635:Fructosediphosphates; D006598:Hexosediphosphates; D010728:Phosphoenolpyruvate; D014302:Trinitrobenzenesulfonic Acid; D002262:Carboxy-Lyases; D010730:Phosphoenolpyruvate Carboxylase; C029063:fructose-1,6-diphosphate; D000645:Ammonium Sulfate