Explain the logic of these pathway regulations:

a. phosphofructokinase, not hexokinase, is the main regulation site of glycolysis.
b. succinylcoa inhibits the entry of acetyl coa into the citric acid cycle.
c. nadh inhibits pyruvate dehydrogenase.
d. citrate inhibits the citric acid cycle and activates acetyl-coa carboxylase.
e. insulin leads to activation of glycogen synthase.

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Explanation:

A Phosphofructokinase is the most important control element in the mammalian glycolytic pathway (Figure 16.16). High levels of ATP allosterically inhibit the enzyme in the liver (a 340-kd tetramer), thus lowering its affinity for fructose 6-phosphate. A high concentration of ATP converts the hyperbolic binding curve of fructose 6-phosphate into a sigmoidal one . ATP elicits this effect by binding to a specific regulatory site that is distinct from the catalytic site. AMP reverses the inhibitory action of ATP, and so the activity of the enzyme increases when the ATP/AMP ratio is lowered. In other words, glycolysis is stimulated as the energy charge falls. A fall in pH also inhibits phosphofructokinase activity. The inhibition of phosphofructokinase by H+ prevents excessive formation of lactic acid and a precipitous drop in blood pH (acidosis).

Phosphofructokinase is the most prominent regulatory enzyme in glycolysis, but it is not the only one. Hexokinase, the enzyme catalyzing the first step of glycolysis, is inhibited by its product, glucose 6-phosphate. High concentrations of this molecule signal that the cell no longer requires glucose for energy, for storage in the form of glycogen, or as a source of biosynthetic precursors, and the glucose will be left in the blood. For example, when phosphofructokinase is inactive, the concentration of fructose 6-phosphate rises. In turn, the level of glucose 6-phosphate rises because it is in equilibrium with fructose 6-phosphate. Hence, the inhibition of phosphofructokinase leads to the inhibition of hexokinase.

Why is phosphofructokinase rather than hexokinase the pacemaker of glycolysis? The reason becomes evident on noting that glucose 6-phosphate is not solely a glycolytic intermediate. Glucose 6-phosphate can also be converted into glycogen or it can be oxidized by the pentose phosphate pathway  to form NADPH. The first irreversible reaction unique to the glycolytic pathway, the committed step, , is the phosphorylation of fructose 6-phosphate to fructose 1,6-bisphosphate. Thus, it is highly appropriate for phosphofructokinase to be the primary control site in glycolysis. In general, the enzyme catalyzing the committed step in a metabolic sequence is the most important control element in the pathway.

B A second control site in the citric acid cycle is α-ketoglutarate dehydrogenase. Some aspects of this enzyme's control are like those of the pyruvate dehydrogenase complex, as might be expected from the homology of the two enzymes. α-Ketoglutarate dehydrogenase is inhibited by succinyl CoA and NADH, the products of the reaction that it catalyzes. In addition, α-ketoglutarate dehydrogenase is inhibited by a high energy charge. Thus, the rate of the cycle is reduced when the cell has a high level of ATP.   The enzyme α-ketoglutarate dehydrogenase is another important catalyst in the fourth step of the cycle where α-ketoglutarate also loses a carbon and combines with Coenzyme A to form succinyl CoA. The two products of the reaction, succinyl CoA and NADH, both work as inhibitors at large concentrations.

C The pyruvate dehydrogenase complex catalyzes an irreversible reaction that is the entry point of pyruvate into the TCA cycle (see below) and is under complex regulation by allosteric and covalent modification of the pyruvate dehydrogenase component of the complex. The end products of the overall reaction (NADH and acetyl-CoA) are potent allosteric inhibitors of the pyruvate dehydrogenase component of the complex.

D Allosteric Regulation : The active dephospho- form of acetyl-CoA carboxylase is regulated by citrate.  Stimulation by citrate assures FAS when 2-carbon units are plentiful.

Citrate is used for feedback inhibition, as it inhibits phosphofructokinase, an enzyme involved in glycolysis that catalyses formation of fructose 1,6-bisphosphate, a precursor of pyruvate. This prevents a constant high rate of flux when there is an accumulation of citrate and a decrease in substrate for the enzyme.

E Insulin stimulates the liver to store glucose in the form of glycogen. A large fraction of glucose absorbed from the small intestine is immediately taken up by hepatocytes, which convert it into the storage polymer glycogen.

Insulin has several effects in liver which stimulate glycogen synthesis. First, it activates the enzyme hexokinase, which phosphorylates glucose, trapping it within the cell. Coincidently, insulin acts to inhibit the activity of glucose-6-phosphatase. Insulin also activates several of the enzymes that are directly involved in glycogen synthesis, including phosphofructokinase and glycogen synthase.

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