Gluconeogenesis pyruvate-phosphoenolpyruvate

OVERSATURATION OXALOACETATE DECARBOXYLASE Oxaloacetate, synthesis in gluconeogenesis, PYRUVATE CARBOXYLASE PHOSPHOENOLPYRUVATE CARBOXYKI-NASE (PYROPHOSPHATE)... 

The first "roadblock" to overcome in the synthesis of glucose from pyruvate is the irreversible conversion in glycolysis of pyruvate to phosphoenolpyruvate (PEP) by pyruvate kinase. In gluconeogenesis, pyruvate is first carboxylated by pyruvate carboxylase to oxaloacetate (OAA), which is then converted to PEP by the action of PEP-carboxykinase (Figure 10.3). 

Figure 21-1. Interconversion of phosphoenolpyruvate (PEP) and pymvate. The conversion of PEP to pyruvate is thermodynamically irreversible in the cell. To convert pyruvate back to PEP for gluconeogenesis, pyruvate must enter the mitochondrion, be carboxylated to oxaloacetate (OAA), and reduced to malate. After exiting the mitochondrion, malate is oxidized back to OAA and converted to PEP by the action of phosphoenolpyruvate carboxykinase.

Oxaloacetate is an intermediate of many metabolic pathways. It also plays a role in the malate-aspartate shuttle, which transfers high energy electrons into mitochondria. Citrate is formed by the condensation of oxaloacetate with acetyl CoA. A transamination reaction transfers an amino group from an amino acid to an a-keto acid. Transfer of the amino group from aspartate to a-ketoglutarate forms oxaloacetate and glutamate. In gluconeogenesis, pyruvate is carboxylated in mitochondria to form oxaloacetate. After transfer to the cytosol, the enzyme phosphoenolpyruvate carboxykinase catalyses the conversion of oxaloacetate to phosphoenolpyruvate. 

A problem for gluconeogenesis is that pyruvate carboxylase, which produces oxaloacetate from pyravate, is present in the mitochondria but phosphoenolpyruvate carboxylase, at least in human liver, is present in the cytosol. For reasons given in Chapter 9, oxaloacetate cannot cross the mitochondrial membrane and so a transporter is not present in any cells. Hence, oxaloacetate is converted to phosphoenolpyruvate which is transported across the membrane (Figure 6.25). 

Figure 6.25 The intracellular location of the gluconeogenic enzymes. The gluconeogenic enzymes are located in the cytosol, except for pyruvate carboxylase which is always present within the mitochondria phosphoenolpyruvate carboxykinase is cytoplasmic in some species including humans. Consequently phosphoenolpyruvate must be transported across the inner mitochondrial enzyme by a transporter molecule in order for gluconeogenesis to take place.

Figure 6-7. Conversion of mitochondrial pyruvate to cytosolic phosphoenolpyruvate to initiate gluconeogenesis. Oxaloacetate cannot pass across the inner mitochondrial membrane, so it is reduced to malate, which can do so.

Gluconeogenesis Consumes ATP Conversion of Pyruvate to Phosphoenolpyruvate Requires Two High Energy Phosphates Conversion of Phosphoenolpyruvate to Fructose-1,6-bisphosphate Uses the Same Enzymes as Glycolysis... 

Pathway of C02 in Gluconeogenesis In the first bypass step of gluconeogenesis, the conversion of pyruvate to phosphoenolpyruvate (PEP), pyruvate is carboxylated by pyruvate carboxylase to oxaloacetate, which is subsequently decarboxylated to PEP by PEP carboxykinase (Chapter 14). Because the addition of C02 is directly followed by the loss of C02, you might expect that in tracer experiments, the 14C of 14C02 would not be incorporated into PEP, glucose, or any intermediates in gluconeogenesis. 

Figure 18.8 Gluconeogenesis pathway in the liver. PC is pyruvate carboxylase PEPCK is phosphoenolpyruvate carboxykinase. (Reproduced by permission from ViduesJ, Sovik O. Gluconeogenesis in infancy and childood. Acta Paediatr Scand 65 307-312, 1976.)...

Step A, the conversion of pyruvate to phosphoenolpyruvate, is accomplished by a circuitous process commencing with pyruvate entering the mitochondrion, which for gluconeogenesis to occur must be in a high-energy state. Under these conditions, the mitochondrial enzyme pyruvate carboxylase catalyzes the conversion of pyruvate to oxaloacetate-. 

Biotin acts to induce glucokinase, phosphofructokinase, and pyruvate kinase (key enzymes of glycolysis), phosphoenolpyruvate carboxykinase (a key enzyme of gluconeogenesis), and holocarboxylase synthetase, acting via a cell-surface receptor linked to formation of cGMP and increased activity of RNA polymerase. The activity of holocarboxylase synthetase (Section 11.2.2) falls in experimental biotin deficiency and increases with a parallel increase in... 

The first step in gluconeogenesis is the carboxylation of pyruvate to form oxaloacetate at the expense of a molecule of ATP. Then, oxaloacetate is decarboxylated and phosphorylated to yield phosphoenolpyruvate, at the expense of the high... 

Finally, oxaloacetate is simultaneously decarboxylated andphosphorylated by phosphoenolpyruvate carboxykinase in the cytosol. The CO2 that vv as added to pyruvate by pyruvate carboxylase comes off in this step. Recall that, in glycolysis, the presence of a phosphoryl group traps the unstable enol isomer of pyruvate as phosphoenolpyruvate (Section 16.1.7). In gluconeogenesis, the formation of the unstable enol is driven by decarboxylation—the oxidation of the carboxylic acid to CO2 —and trapped by the addition of a phosphate to carbon 2 from GTP. The two-step pathway... 

A third fate of pyruvate is its carboxylation to oxaloacetate inside mitochondria, the first step in gluconeogenesis. This reaction and the subsequent conversion of oxaloacetate into phosphoenolpyruvate bypass an irreversible step of glycolysis and hence enable glucose to be synthesized from pyruvate. The carboxylation of pyruvate is also important for replenishing intermediates of the citric acid cycle. Acetyl CoA activates pyruvate carboxylase, enhancing the synthesis of oxaloacetate, when the citric acid cycle is slowed by a paucity of this intermediate. 

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