Effects of adenosine phosphates and nicotinamide nucleotides on pyruvate carboxylase from baker's yeast.
1. Pyruvate carboxylase from baker's yeast is inhibited by ADP, AMP and adenosine at pH8.0 in the presence of magnesium chloride concentrations equal to or higher than the ATP concentration. The adenine moiety is essential for the inhibitory effect. 2. In the absence of acetyl-CoA (an allosteric activator) ADP, AMP and adenosine are competitive inhibitors with respect to ATP. In the presence of acetyl-CoA, besides the effect with respect to ATP, AMP competes with acetyl-CoA, whereas ADP and adenosine are non-competitive inhibitors with respect to the activator. 3. Pyruvate carboxylase is inhibited by NADH. The inhibition is competitive with respect to acetyl-CoA and specific with respect to NADH, since NAD(+), NADP(+) and NADPH do not affect the enzyme activity. In the absence of acetyl-CoA, NAD(+), NADH, NADP(+) and NADPH do not inhibit pyruvate carboxylase. 4. Pyruvate carboxylase is inhibited by ADP, AMP and NADH at pH6.5, in the presence of 12mm-Mg(2+), 0.75mm-Mn(2+) and 0.5mm-ATP, medium conditions similar to those existing inside the yeast cell. The ADP and NADH effects are consistent with a regulation of enzyme activity by the intracellular [ATP]/[ADP] ratio and secondarily by NADH concentration. These mechanisms would supplement the already known control of yeast pyruvate carboxylase by acetyl-CoA and l-aspartate. Inhibition by AMP is less marked and its physiological role is perhaps limited.
D000225:Adenine; D000227:Adenine Nucleotides; D000255:Adenosine Triphosphate; D055598:Chemical Phenomena; D002621:Chemistry; D003065:Coenzyme A; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008025:Ligases / Q000037:antagonists & inhibitors; D008274:Magnesium; D008345:Manganese; D009243:NAD; D009249:NADP; D009705:Nucleosides; D010071:Oxaloacetates; D011773:Pyruvates; D012440:Saccharomyces / Q000201:enzymology*
D000227:Adenine Nucleotides; D009705:Nucleosides; D010071:Oxaloacetates; D011773:Pyruvates; D009243:NAD; D008345:Manganese; D009249:NADP; D000255:Adenosine Triphosphate; D008025:Ligases; D008274:Magnesium; D000225:Adenine; D003065:Coenzyme A