On the mechanism of inhibition of gluconeogenesis and ureagenesis by sodium benzoate.
Synthesis of glucose from lactate and generation of urea from ammonia were inhibited when sodium benzoate was added to suspensions of rat hepatocytes. Assays with isolated mitochondria suggested pyruvate carboxylase and the N-acetyl-L-glutamate (NAG)-dependent carbamoylphosphate synthetase (CPS-I) as potential sites of inhibition for both pathways, owing to a shared dependency on aspartate efflux from the mitochondria and its subsequent conversion to oxaloacetate in the cytosol. Assays with isolated hepatocytes indicated inhibition to be initiated by accumulation of benzoyl CoA with a resultant depletion of free CoA and acetyl CoA. Measurements of adenine nucleotides showed that benzoate metabolism did not sufficiently alter energy status to account for the observed inhibition. Consistent with these interpretations, acceleration of the conversion of benzoyl CoA to hippurate by the addition of glycine restored the levels of free CoA and acetyl CoA and the rates of gluconeogenesis and ureagenesis. Reduction of the levels of aspartate and glutamate, presumably by interference with the anapleurotic function of pyruvate carboxylase, most likely accounted for inhibition of gluconeogenesis by benzoate. Whether reduced flux through the urea cycle also contributed to inhibition of gluconeogenesis (by diminishing cytosolic conversion of aspartate to oxaloacetate) requires further study. Depression of glutamate and acetyl CoA to levels at or below the Km for NAG synthetase probably accounted for the observed inhibition of ureagenesis. Rates of urea production were observed to vary with changes in the levels of NAG, suggesting NAG-dependent CPS-I to be the primary site of inhibition of ureagenesis by benzoate.
Department of Biochemistry and Biophysics, University of Rhode Island, Kingston 02881.
D M
DM
Cyr
S G
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Egan
C M
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Brini
G C
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Tremblay
D000105:Acetyl Coenzyme A / Q000378:metabolism; D000227:Adenine Nucleotides / Q000378:metabolism; D000818:Animals; D001224:Aspartic Acid / Q000378:metabolism; D001565:Benzoates / Q000494:pharmacology*; D019817:Benzoic Acid; D001639:Bicarbonates / Q000378:metabolism; D002222:Carbamoyl-Phosphate Synthase (Ammonia) / Q000378:metabolism; D002478:Cells, Cultured / Q000187:drug effects; D004789:Enzyme Activation; D005943:Gluconeogenesis / Q000187:drug effects*; D005971:Glutamates / Q000378:metabolism; D018698:Glutamic Acid; D007700:Kinetics; D008099:Liver / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D008930:Mitochondria, Liver / Q000201:enzymology; D009963:Orotic Acid / Q000378:metabolism; D011766:Pyruvate Carboxylase / Q000378:metabolism; D051381:Rats; D012964:Sodium / Q000378:metabolism; D017693:Sodium Bicarbonate; D014508:Urea / Q000378:metabolism*
D000227:Adenine Nucleotides; D001565:Benzoates; D001639:Bicarbonates; D005971:Glutamates; D001224:Aspartic Acid; D018698:Glutamic Acid; D009963:Orotic Acid; D000105:Acetyl Coenzyme A; D017693:Sodium Bicarbonate; D019817:Benzoic Acid; D014508:Urea; D012964:Sodium; D002222:Carbamoyl-Phosphate Synthase (Ammonia); D011766:Pyruvate Carboxylase; C016195:N-acetylglutamic acid