Regulation of Glycolysis

Regulation in muscles
Glycolysis in muscle is regulated to meet the need for ATP.
Glycolysis in skeletal muscle provides ATP primarily to power
contraction. Consequently, the primary control of muscle
glycolysis is the energy charge of the cell—the ratio of ATP to
AMP. Let us examine how each of the key regulatory enzymes
responds to changes in the amounts of ATP and AMP present in
the cell.
Phosphofructokinase
Hexokinase
Pyruvate kinase
All of these enzymes in the muscles are activated by low ATP
and inhibited by high ATP concentration.

Regulation in Liver
Phosphofructokinase.
Regulation with respect to ATP is the same in the liver as in
muscle. Low pH is not a metabolic signal for the liver enzyme,
because lactate is not normally produced in the liver. Indeed, as
we will see, lactate is converted into glucose in the liver.
Glycolysis also furnishes carbon skeletons for biosyntheses, and
so a signal indicating whether building blocks are abundant or
scarce should also regulate phosphofructokinase. In the liver,
phosphofructokinase is inhibited by citrate, an early
intermediate in the citric acid cycle. A high level of citrate in the
cytoplasm means that biosynthetic precursors are abundant, and
so there is no need to degrade additional glucose for this
purpose. Citrate inhibits phosphofructokinase by enhancing
the inhibitory effect of ATP. One means by which glycolysis in
the liver responds to changes in blood glucose is through the
signal molecule fructose 2,6-bisphosphate (F-2,6-BP), a potent
activator of phosphofructokinas. In the liver, the concentration
of fructose 6-phosphate rises when blood-glucose concentration
is high, and the abundance of fructose 6-phosphate accelerates
the synthesis of F-2,6-BP. Hence, an abundance of fructose 6-
phosphate leads to a higher concentration of F-2,6-BP. The
binding of fructose 2,6-bisphosphate increases the affinity of
phosphofructokinase for fructose 6-phosphate and
diminishes the inhibitory effect of ATP. Glycolysis is thus
accelerated when glucose is abundant. Such a process is called
feedforward stimulation. We will turn to the synthesis and
degradation of this important regulatory molecule after we have
considered gluconeogenesis.
Hexokinase. The hexokinase reaction in the liver is controlled
as in the muscle. However, the liver, in keeping with its role as
monitor of blood glucose levels, possesses another specialized
isozyme of hexokinase, called glucokinase, which is not
inhibited by glucose 6-phosphate. Glucokinase phosphorylates
glucose only when glucose is abundant because the affinity of
glucokinase for glucose is about 50-fold lower than that of
hexokinase. The role of glucokinase is to provide glucose 6-
phosphate for the synthesis of glycogen and for the formation
of fatty acids. The low affinity of glucokinase for glucose in the
liver gives the brain and muscles first call on glucose when its
supply is limited, and it ensures that glucose will not be wasted
when it is abundant. Glucokinase is also present in the β cells of
the pancreas, where the increased formation of glucose 6-
phosphate by glucokinase when blood-glucose levels are
elevated leads to the secretion of the hormone insulin. Insulin
signals the need to remove glucose from the blood for storage as
glycogen or conversion into fat.
Pyruvate kinase
When the blood-glucose level is low, the glucagon-triggered
cyclic AMP cascade leads to the phosphorylation of pyruvate
kinase, which diminishes its activity. This hormone-triggered
phosphorylation prevents the liver from consuming glucose
when it is more urgently needed by the brain and muscle.
Fructose and galactose are converted into glycolytic
intermediates
Although glucose is the most widely used monosaccharide,
others also are important fuels. Let us consider how two
abundant sugars—fructose and galactose—can be funneled
into the glycolytic pathway. There are no catabolic pathways
for metabolizing fructose or galactose, and so the strategy is to
convert these sugars into a metabolite of glucose. Fructose can
take one of two pathways to enter the glycolytic pathway. Much
of the ingested fructose is metabolized by the liver, using the
fructose 1-phosphate pathway.

The first step is the phosphorylation of fructose to fructose 1-

phosphate by fructokinase. Fructose 1-phosphate is then split
into glyceraldehyde and phosphate by Aldolase.
Glyceraldehyde is then phosphorylated to glyceraldehyde 3-
phosphate, a glycolytic intermediate, by triose kinase. In other
tissues, fructose can be phosphorylated to fructose 6-phosphate
by hexokinase.
Galactose is converted into glucose 6-phosphate in four steps.
The first reaction in the galactose–glucose interconversion
pathway is the phosphorylation of galactose to galactose 1-
phosphate by galactokinase.

Galactose 1-phosphate then acquires a uridyl group from uridine

diphosphate glucose (UDP-glucose), an activated intermediate
in the synthesis of carbohydrates.
The products of this reaction, which is catalyzed by galactose 1-
phosphate uridyl transferase, are UDP-galactose and glucose 1-
phosphate. The galactose moiety of UDP-galactose is then
epimerized to glucose. The configuration of the hydroxyl group
at carbon 4 is inverted by UDP-galactose 4-epimerase.
Finally, glucose 1-phosphate, formed from galactose, is
isomerized to glucose 6-phosphate by phosphoglucomutase. We
shall return to this reaction when we consider the synthesis and
degradation of glycogen, which proceeds through glucose 1-
phosphate,
Many adults are intolerant of milk because they are deficient
in lactase
Many adults are unable to metabolize the milk sugar lactose and
experience gastrointestinal disturbances if they drink milk.
Lactose intolerance, or hypolactasia, is most commonly caused
by a deficiency of the enzyme lactase, which cleaves lactose into
glucose and galactose.
“Deficiency” is not quite the appropriate term, because a
decrease in lactase is normal in the course of development in all
mammals. As children are weaned and milk becomes less
prominent in their diets, lactase activity normally declines to
about 5% to 10% of the level at birth. This decrease is not as
pronounced with some groups of people, most notably Northern
Europeans, and people from these groups can continue to ingest
milk without gastrointestinal difficulties. With the appearance of
milk-producing domesticated animals, an adult with active
lactase would have a selective advantage in being able to
consume calories from the readily available milk. Because dairy
farming originated only about 10,000 years ago, the
evolutionary selective pressure on lactase persistence must have
been substantial, attesting to the biochemical value of being able
to use milk as an energy source into adulthood.
What happens to the lactose in the intestine of a lactase-deficient
person? The lactose is a good energy source for microorganisms
in the colon, and they ferment it to lactic acid while generating
methane (CH4) and hydrogen gas (H2). The gas produced
creates the uncomfortable feeling of gut distension and the
annoying problem of flatulence. The lactate produced by the
microorganisms is osmotically active and draws water into the
intestine, as does any undigested lactose, resulting in diarrhea. If
severe enough, the gas and diarrhea hinder the absorption of
other nutrients such as fats and proteins. The simplest treatment
is to avoid the consumption of products containing much
lactose. Alternatively, the enzyme lactase can be ingested with
milk products.
Galactose is highly toxic if the transferase is missing
Less common than lactose intolerance are disorders that
interfere with the metabolism of galactose. The disruption of
galactose metabolism is referred to as galactosemia. The most
common form, called classic galactosemia, is an inherited
deficiency in galactose 1-phosphate uridyl transferase activity.
Afflicted infants fail to thrive. They vomit or have diarrhea after
consuming milk, and enlargement of the liver and jaundice are
common, sometimes progressing to cirrhosis. Cataracts will
form, and lethargy and retarded mental development also are
common. The blood-galactose level is markedly elevated, and
galactose is found in the urine. The absence of the transferase in
red blood cells is a definitive
diagnostic criterion.
The most common treatment is to remove galactose (and
lactose) fromthe diet. An enigma of galactosemia is that,
although elimination of galactose from the diet prevents liver
disease and cataract development, the majority of patients still
suffer from central nervous system malfunction, most commonly
a delayed acquisition of language skills. Female patients also
display ovarian failure. Cataract formation is better understood.
A cataract is the clouding of thenormally clear lens of the eye. If
the transferase is not active in the lens of the eye, the presence of
aldose reductase causes the accumulating galactose to be
reduced to galactitol.
Galactitol is osmotically active, and water will diffuse into the
lens, instigating the formation of cataracts. In fact, there is a
high incidence of cataract formation with age in populations that
consume substantial amounts of milk into adulthood.

A family of transporters enables glucose to enter and leave

animal cells
Several glucose transporters mediate the thermodynamically
downhillmovement of glucose across the plasma membranes of
animal cells. Eachmember of this protein family, named GLUT1
to GLUT5, consists of asingle polypeptide chain about 500
residues long.
The members of this family have distinctive roles:
1. GLUT1 and GLUT3, present in nearly all mammalian cells,
are responsible for basal glucose uptake, GLUT1 and GLUT3
continually transport glucose into cells at an essentially constant
rate.
2. GLUT2, present in liver and pancreatic β cells, glucose enters
these tissues at a biologically significant rate only when there is
much glucose in the blood. The pancreas can sense the glucose
level and accordingly adjust the rate of insulin secretion.
3. GLUT4, transports glucose into muscle and fat cells. The
number of GLUT4 transporters in the plasma membrane
increases rapidly in the presence of insulin, which signals the
fed state. Hence, insulin promotes the uptake of glucose by
muscle and fat. Endurance exercise training increases the
amount of this transporter present
in muscle membranes.
4. GLUT5, present in the small intestine, functions primarily as
a fructose transporter.
Cancer and exercise training affect glycolysis in a similar
fashion
Tumors have been known for decades to display enhanced rates
of glucose uptake and glycolysis. Indeed, rapidly growing tumor
cells will metabolize glucose to lactate even in the presence of
oxygen, a process called aerobic glycolysis or the “Warburg
effect,” after Otto Warburg, the biochemist who first noted this
characteristic of cancer cells in the 1920s. In fact, tumors with a
high glucose uptake are particularly aggressive, and the cancer is
likely to have a poor prognosis. Positron emission tomography
(PET) and computer-aided tomography (CAT), can easily
visualizes tumors.
What selective advantage does aerobic glycolysis offer the
tumor over the energetically more efficient oxidative
phosphorylation? Research is being actively pursued to answer
the question, but we can speculate on the benefits. First, aerobic
glycolysis generates lactic acid that is then secreted.
Acidification of the tumor environment has been shown to
facilitate tumor invasion and inhibit the immune system from
attacking the tumor. Second, the increased uptake of glucose and
formation of glucose 6-phosphate provides substrates for
another metabolic pathway, the pentose phosphate that
generates biosynthetic reducing power. Moreover, the pentose
phosphate pathway, in cooperation with glycolysis, produces
precursors for biomolecules necessary for growth, such as
nucleotides. Finally, cancer cells grow more rapidly than the
blood vessels that nourish them; thus, as solid tumors grow, the
oxygen concentration in their environment falls. In other words,
they begin to experience hypoxia, a deficiency of oxygen. The
use of aerobic glycolysis reduces the dependence of cell growth
on oxygen. Hypoxia itself enhances tumor growth by activating
a transcription factor, hypoxia-inducible transcription factor
(HIF-1). HIF-1 increases the expression of most glycolytic
enzymes and the glucose transporters GLUT1 and GLUT3.
These adaptations by the cancer cells enable a tumor to survive
until blood vessels can grow. HIF-1 also increases the
expression of signal molecules, such as vascular endothelial
growth factor (VEGF), that facilitate the growth of blood vessels
that will provide nutrients to the cells. Without new blood
vessels, a tumor would cease to grow and either die or remain
harmlessly small. Efforts are underway to develop drugs that
inhibit the growth of blood vessels in tumors.