The effects of ammonium chloride and bicarbonate on the activity of glutaminase in isolated liver mitochondria.
1. Glutamine hydrolysis in liver mitochondria was studied by measuring the production of glutamate under conditions where this compound could not be further metabolized. 2. Glutaminase activity in intact mitochondria was very low in the absence of activators. 3. Glutamine hydrolysis was markedly stimulated by NH4Cl and also by HCO3- ions. 4. The stimulation by each of these compounds was much decreased if the mitochondria were uncoupled. 5. Maximum rates of glutamine hydrolysis required the addition of phosphate. A correlation was observed between the activity of glutaminase in the presence of NH4Cl plus HCO3- and the intramitochondrial content of ATP. 6. In disrupted mitochondria, NH4Cl stimulated glutaminase to a much smaller extent than in intact mitochondria. The NH4Cl stimulation in disrupted mitochondria was much increased by the addition of ATP. KHCO3 also stimulated glutaminase activity in disrupted mitochondria, and ATP increased the magnitude of this stimulation. 7. It was concluded that maximum rates of glutaminase activity in liver mitochondria require the presence of phosphate, ATP and either HCO3- or NH4+. A comparison of the results obtained on intact and broken mitochondria indicates that these effectors have a direct effect on the glutaminase enzyme system rather than an indirect effect mediated by changes in transmembrane ion gradients or in the concentrations of intramitochondrial metabolites.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000643:Ammonium Chloride / Q000494:pharmacology*; D000818:Animals; D001639:Bicarbonates / Q000494:pharmacology*; D004734:Energy Metabolism; D005260:Female; D005972:Glutaminase / Q000378:metabolism*; D005973:Glutamine / Q000378:metabolism; D066298:In Vitro Techniques; D008930:Mitochondria, Liver / Q000187:drug effects / Q000201:enzymology*; D051381:Rats
D001639:Bicarbonates; D000643:Ammonium Chloride; D005973:Glutamine; D000255:Adenosine Triphosphate; D005972:Glutaminase