Glucose Can Be Synthesized from Noncarbohydrate Precursors

We now turn to the synthesis of glucose from noncarbohydrate

precursors, a process called gluconeogenesis. Maintaining levels of
glucose is important because the brain depends on glucose as its
primary fuel and red blood cells use glucose as their only fuel. The daily
glucose requirement of the brain in a typical adult human being is about
120 g, which accounts for most of the 160 g of glucose needed daily by
the whole body. The amount of glucose present in body fluids is about
20 g, and that readily available from glycogen is approximately 190 g.
Thus, the direct glucose reserves are sufficient to meet glucose needs for
about a day. Gluconeogenesis is especially important during a longer
period of fasting or starvation. The gluconeogenic pathway converts
pyruvate into glucose. Noncarbohydrate precursors of glucose are first
converted into pyruvate or enter the pathway at later intermediates such
as oxaloacetate and dihydroxyacetone phosphate (Figure 16.24). The
major noncarbohydrate precursors are lactate, amino acids, and
glycerol. Lactate is formed by active skeletal muscle when the rate of
glycolysis exceeds the rate of oxidative metabolism. Lactate is readily
converted into pyruvate by the action of lactate dehydrogenase.
Amino acids are derived from proteins in the diet. The hydrolysis of
triacylglycerols in fat cells yields glycerol and fatty acids. Glycerol may
enter either the gluconeogenic or the glycolytic pathway at
dihydroxyacetone phosphate.
The major site of gluconeogenesis is the liver, with a small amount also
taking place in the kidney. Little gluconeogenesis takes place in the
brain, skeletal muscle, or heart muscle. Rather, gluconeogenesis in the
liver and kidney helps to maintain the glucose level in the blood so that
the brain and muscle can extract sufficient glucose from it to meet their
metabolic demands.

Gluconeogenesis and Glycolysis Are Reciprocally Regulated

Gluconeogenesis and glycolysis are coordinated so that, within a cell,
one pathway is relatively inactive while the other is highly active. If both
sets of reactions were highly active at the same time, the net result
would be the hydrolysis of four nucleoside triphosphates (two ATP
molecules plus two GTP molecules) per reaction cycle. However, the
amounts and activities of the distinctive enzymes of each pathway are
controlled so that both pathways are not highly active at the same time.
The rate of glycolysis is also determined by the concentration of
glucose, and the rate of gluconeogenesis by the concentrations of lactate
and other precursors of glucose. The basic premise of the reciprocal
regulation is that, when energy is needed, glycolysis will predominate.
When there is a surplus of energy, gluconeogenesis will take over.
Energy charge determines whether glycolysis or gluconeogenesis will be
most active The first important regulation site is the interconversion of
fructose 6-phosphate and fructose 1,6-bisphosphate. Consider first a
situation in which energy is needed. In this case, the concentration of
AMP is high. Under this condition, AMP stimulates
phosphofructokinase but inhibits fructose 1,6-bisphosphatase. Thus,
glycolysis is turned on and gluconeogenesis is inhibited. Conversely,
high levels of ATP and citrate indicate that the energy charge is high and
that biosynthetic intermediates are abundant. ATP and citrate inhibit
phosphofructokinase, whereas citrate activates fructose 1,6-
bisphosphatase. Under these conditions, glycolysis is nearly switched off
and gluconeogenesis is promoted.
Why does citrate take part in this regulatory scheme? Citrate reports on
the status of the citric acid cycle, the primary pathway for oxidizing
fuels in the presence of oxygen. High levels of citrate indicate an
energy-rich situation and the presence of precursors for biosynthesis.

Glycolysis and gluconeogenesis are also reciprocally regulated at the

interconversion of phosphoenolpyruvate and pyruvate in the liver. The
glycolytic enzyme pyruvate kinase is inhibited by allosteric effectors
ATP and alanine, which signal that the energy charge is high and that
building blocks are abundant. Conversely, pyruvate carboxylase, which
catalyzes the first step in gluconeogenesis from pyruvate, is inhibited by
ADP. Likewise, ADP inhibits phosphoenolpyruvate carboxykinase.
Pyruvate carboxylase is activated by acetyl CoA, which, like citrate,
indicates that the citric acid cycle is producing energy and biosynthetic
intermediates. Hence, gluconeogenesis is favored when the cell is rich in
biosynthetic
precursors and ATP.

The balance between glycolysis and gluconeogenesis in the liver is

sensitive to blood-glucose concentration
In the liver, rates of glycolysis and gluconeogenesis are adjusted to
maintain blood-glucose levels. The signal molecule fructose 2,6-
bisphosphate strongly stimulates phosphofructokinase (PFK) and
inhibits fructose 1,6-bisphosphatase. When blood glucose is low,
fructose 2,6-bisphosphate loses a phosphoryl group to form fructose 6-
phosphate, which no longer binds to PFK. How is the concentration of
fructose 2,6-bisphosphate controlled to rise and fall with blood-glucose
levels? Two enzymes regulate the concentration of this molecule: one
phosphorylates fructose 6-phosphate and the other dephosphorylates
fructose 2,6-bisphosphate. Fructose 2,6-bisphosphate is formed in a
reaction catalyzed by phosphofructokinase 2 (PFK2), a different enzyme
from phosphofructokinase. Fructose 6-phosphate is formed through the
hydrolysis of fructose 2,6-bisphosphate by a specific phosphatase,
fructose bisphosphatase 2 (FBPase2). The striking finding is that both
PFK2 and FBPase2 are present in a single 55-kd
polypeptide chain (Figure 16.31). This bifunctional enzyme contains an
N-terminal regulatory domain, followed by a kinase domain and a
phosphatase domain.
When glucose is scarce, such as during a night’s fast, a rise in the blood
level of the hormone glucagon triggers a cyclic AMP signal cascade,
leading to the phosphorylation of this bifunctional enzyme by protein
kinase A (Figure 16.32). This covalent modification activates FBPase2
and inhibits PFK2, lowering the level of F-2,6-BP. Gluconeogenesis
predominates. Glucose formed by the liver under these conditions is
essential for the viability of the brain. Glucagon stimulation of protein
kinase A also inactivates pyruvate kinase in the liver Conversely, when
blood-glucose levels are high, such as after a meal, gluconeogenesis is
not needed. Insulin is secreted and initiates a signal pathway that
activates a protein phosphatase, which removes the phosphoryl group
from the bifunctional enzyme. This covalent modification activates
PFK2 and inhibits FBPase2. The resulting rise in the level of F-2,6-BP
accelerates glycolysis. The coordinated control of glycolysis and
gluconeogenesis is facilitated by the location of the kinase and
phosphatase domainson the same polypeptide chain as the regulatory
domain.The hormones insulin and glucagon also regulate the amounts of
essential enzymes. These hormones alter gene expression primarily by
changing the rate of transcription. Insulin levels rise subsequent to
eating, when there is plenty of glucose for glycolysis. To encourage
glycolysis, insulin stimulates the expression of phosphofructokinase,
pyruvate kinase, and the
bifunctional enzyme that makes and degrades F-2,6-BP. Glucagon rises
during fasting, when gluconeogenesis is needed to replace scarce
glucose. To encourage gluconeogenesis, glucagon inhibits the
expression of the three regulated glycolytic enzymes and stimulates
instead the production of two key gluconeogenic enzymes,
phosphoenolpyruvate carboxykinase and
fructose 1,6-bisphosphatase.